Aminoalkyl glucosaminide phosphate compounds and their use as adjuvants and immunoeffectors

ABSTRACT

Aminoalkyl glucosaminide phosphate (AGP) compounds that are adjuvants and immunoeffectors are described and claimed. The compounds have a 2-deoxy-2-amino glucose in glycosidic linkage with an aminoalkyl (aglycon) group. Compounds are phosphorylated at the 4 or 6 carbon on the glucosaminide ring and comprise three 3-alkanoyloxyalkanoyl residues. The compounds augment antibody production in immunized animals as well as stimulate cytokine production and activate macrophages. Methods for using the compounds as adjuvants and immunoeffectors are also disclosed.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation-in-part of U.S. application Ser. No.08/853,826, filed May 8, 1997 (which application is hereby incorporatedby reference in its entirety).

TECHNICAL FIELD

This invention relates generally to aminoalkyl glucosaminide phosphate(AGP) compounds which have activity as adjuvants and immunoeffectors,and methods and compositions related thereto.

BACKGROUND OF THE INVENTION

Humoral immunity and cell-mediated immunity are the two major branchesof the mammalian immune response. Humoral immunity involves thegeneration of antibodies to foreign antigens. Antibodies are produced byB-lymphocytes. Cell-mediated immunity involves the activation ofT-lymphocytes which either act upon infected cells bearing foreignantigens or stimulate other cells to act upon infected cells. Bothbranches of the mammalian immune system are important in fightingdisease. Humoral immunity is the major line of defense against bacterialpathogens. In the case of viral disease, the induction of cytotoxic Tlymphocytes (CTLs) appears to be crucial for protective immunity. Aneffective vaccine stimulates both branches of the immune system toprotect against disease.

Vaccines present foreign antigens from disease causing agents to a hostso that the host can mount a protective immune response. Often vaccineantigens are killed or attenuated forms of the microbes which cause thedisease. The presence of non-essential components and antigens in thesekilled or attenuated vaccines has encouraged considerable efforts torefine vaccine components including developing well-defined syntheticantigens using chemical and recombinant techniques. The refinement andsimplification of microbial vaccines, however, has led to a concomitantloss in potency. Low-molecular weight synthetic antigens, though devoidof potentially harmful contaminants, are themselves not veryimmunogenic. These observations have led investigators to add adjuvantsto vaccine compositions to potentiate the activity of the refinedvaccine components.

Presently, the only adjuvant licensed for human use in the United Statesis alum, a group of aluminum salts (e.g., aluminum hydroxide, aluminumphosphate) in which vaccine antigens are formulated. Particulatecarriers like alum serve to promote the uptake, processing andpresentation of soluble antigens by macrophages. Alum, however, is notwithout side-effects and enhances humoral (antibody) immunity only.

An effective adjuvant potentiates both a humoral and cellular immuneresponse in vaccinated animals. Further, an adjuvant must enhance ahost's natural immune response and not aggravate the host system. Awell-defined synthetic adjuvant free from extraneous matter which isstable and easy to manufacture would provide these qualities. Compoundsthat have been prepared and tested for adjuvanticity (Shimizu et al.1985, Bulusu et al. 1992, Ikeda et al. 1993, Shimizu et al. 1994,Shimizu et al. 1995, Miyajima et al. 1996), however, often display toxicproperties, are unstable and/or have unsubstantial immunostimulatoryeffects.

The discovery and development of effective adjuvants is essential forimproving the efficacy and safety of existing vaccines. Adjuvants impartsynthetic peptides and carbohydrate antigens with sufficientimmunogenicity to insure the success of the synthetic vaccine approach.There remains a need for new compounds having potent immunomodulatingeffects.

SUMMARY OF THE INVENTION

The compounds of the subject invention are aminoalkyl glucosaminidephosphate compounds (AGPs) which are adjuvants and immunoeffectors. Anaminoalkyl (aglycon) group is glycosidically linked to a2-deoxy-2-amino-α-D-glucopyranose (glucosaminide) to form the basicstructure of the claimed molecules. The compounds are phosphorylated atthe 4 or 6 carbon on the glucosaminide ring. Further, the compoundspossess three 3-alkanoyloxyalkanoyl residues.

The compounds of the subject invention are immunoeffector moleculesaugmenting antibody production in immunized animals, stimulatingcytokine production and activating macrophages. In accordance with the-subject invention, methods for using these compounds as adjuvants andimmunoeffectors are disclosed.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the subject invention are adjuvant and immunoeffectormolecules which are aminoalkyl glucosaminide phosphates (AGPs). Thecompounds comprise a 2-deoxy-2-amino-α-D-glucopyranose (glucosaminide)in glycosidic linkage with an aminoalkyl (aglycon) group. Compounds arephosphorylated at the 4 or 6 carbon on the glucosaminide ring and havethree alkanoyloxyalkanoyl residues. The compounds of the subjectinvention are described generally by Formula I,

wherein X represents an oxygen or sulfur atom in either the axial orequitorial position, Y represents an oxygen atom or NH group, “n”, “m”,“p” and “q” are integers from 0 to 6, R₁, R₂, and R₃ represent normalfatty acyl residues having 1 to 20 carbon atoms and where one of R₁, R₂or R₃ is optionally hydrogen, R₄ and R₅ are hydrogen or methyl, R₆ andR₇ are hydrogen, hydroxy, alkoxy, phosphono, phosphonooxy, sulfo,sulfooxy, amino, mercapto, cyano, nitro, formyl or carboxy and estersand amides thereof; R₈ and R₉ are phosphono or hydrogen. Theconfiguration of the 3′ stereogenic centers to which the normal fattyacyl residues are attached is R or S, but preferably R. Thestereochemistry of the carbon atoms to which R₄ or R₅ are attached canbe R or S. All stereoisomers, both enantiomers and diastereomers, andmixtures thereof, are considered to fall within the scope of the subjectinvention.

The heteroatom X of the compounds of the subject invention can be oxygenor sulfur. In a preferred embodiment, X is oxygen and typically in theequitorial position. Although the stability of the molecules could beeffected by a substitution at X, the immunomodulating activity ofmolecules with these substitutions is not expected to change.

The number of carbon atoms between heteroatom X and the aglycon nitrogenatom is determined by variables “n” and “m”. Variables “n” and “m” canbe integers from 0 to 6. In a preferred embodiment, the total number ofcarbon atoms between heteroatom X and the aglycon nitrogen atom is fromabout 2 to about 6 and most preferably from about 2 to about 4.

The compounds of the subject invention are aminoalkyl glucosaminidecompounds which are phosphorylated. Compounds can be phosphorylated atposition 4 or 6 (R₈ or R₉) on the glucosaminide ring and are mosteffective if phosphorylated on at least one of these positions. In apreferred embodiment, R₈ is phosphono and R₉ is hydrogen.

In one embodiment, the compounds of the subject invention arehexaacylated, that is they contain a total of six fatty acid residues.The aminoalkyl glucosaminide moiety is acylated at the 2-amino and3-hydroxyl groups of the glucosaminide unit and at the amino group ofthe aglycon unit with 3-hydroxyalkanoyl residues. In Formula I, thesethree positions are acylated with 3-hydroxytetradecanoyl moieties. The3-hydroxytetradecanoyl residues are, in turn, substituted with normalfatty acids (R₁-R₃), providing three 3-n-alkanoyloxytetradecanoylresidues or six fatty acid groups in total.

In another embodiment, the compounds of the subject invention arepentaacylated, that is they contain a total of five fatty acid residues.More specifically, the 3-hydroxytetradecanoyl residues of Formula I aresubstituted with normal fatty acids at two of the three R₁, R₂ and R₃positions, with the third R₁, R₂ or R₃ position being hydrogen. In otherwords, at least one of —OR₁, —OR₂ or —OR₃ is hydroxyl.

The chain length of normal fatty acids R₁-R₃ can be from 1 to about 20,and typically from about 7 to about 16 carbons. Preferably, R₁-R₃ arefrom about 9 to about 14 carbons. The chain lengths of these normalfatty acids can be the same or different. Although, only normal fattyacids are described, it is expected that unsaturated fatty acids (i.e.fatty acid moieties having double or triple bonds) substituted at R₁-R₃on the compounds of the subject invention would produce biologicallyactive molecules. Further, slight modifications in the chain length ofthe 3-hydroxyalkanoyl residues are not expected to dramatically effectbiological activity.

The compounds of the subject invention are adjuvants and immunoeffectorswhich enhance the generation of antibody in immunized animals, stimulatethe production of cytokines and stimulate a cell-mediated immuneresponse including a cytotoxic T-lymphocyte response. In methods foreffecting the immune response of an individual, the compounds of thesubject invention can be formulated with a pharmaceutically acceptablecarrier for injection or ingestion. As used herein, “pharmaceuticallyacceptable carrier” means a medium which does not interfere with theimmunomodulatory activity of the active ingredient and is not toxic tothe patient to whom it is administered. Pharmaceutically acceptablecarriers include oil-in-water or water-in-oil emulsions, aqueouscompositions, liposomes, microbeads and microsomes. For example, thecarrier may be a microsphere or microparticle having a compound of thisinvention within the matrix of the sphere or particle or adsorbed on thesurface of the sphere or particle. The carrier may also be an aqueoussolution or micellar dispersion containing triethylamine,triethanolamine or other agent that renders the formulation alkaline innature, or a suspension containing aluminum hydroxide, calciumhydroxide, calcium phosphate or tyrosine adsorbate.

Formulations of the compounds of the subject invention that can beadministered parenterally, i.e. intraperitoneally, subcutaneously orintramuscularly include the following preferred carriers. Examples ofpreferred carriers for subcutaneous use include a phosphate bufferedsaline (PBS) solution and 0.01-0.1% triethanolamine in USP Water forInjection. Suitable carriers for intramuscular injection include 10% USPethanol, 40% propylene glycol and the balance an acceptable isotonicsolution such as 5% dextrose.

Examples of preferred carriers for intravenous use include 10% USPethanol, 40% USP propylene glycol and the balance USP Water forInjection. Another acceptable carrier includes 10% USP ethanol and USPWater for Injection; yet another acceptable carrier is 0.01-0.1%triethanolamine in USP Water for Injection. Pharmaceutically acceptableparenteral solvents are such as to provide a solution or dispersion maybe filtered through a 5 micron filter without removing the activeingredient.

Examples of carriers for administration via mucosal surfaces depend uponthe particular route. When administered orally, pharmaceutical grades ofmannitol, starch, lactose, magnesium stearate, sodium saccharide,cellulose, magnesium carbonate and the like, with mannitol beingpreferred. When administered intranasally, polyethylene glycol orglycols, sucrose, and/or methylcellulose, and preservatives such asbenzalkonium chloride, EDTA, may be used, with polyethylene glycolsbeing preferred, and when administered by inhalation, suitable carriersare polyethylene glycol or glycols, methylcellulose, dispensing agents,and preservatives, with polyethylene glycols being preferred.

The compounds of the subject invention are administered to an individualin “an effective amount” to effect or enhance the individual's immuneresponse. As used herein, “an effective amount” is that amount whichshows a response over and above the vehicle or negative controls. Theprecise dosage of the compounds of the subject invention to beadministered to a patient will depend upon the particular AGP used, theroute of administration, the pharmaceutical composition, and thepatient. For example, when administered subcutaneously to enhance anantibody response, the amount of AGP used is from 1 to about 250micrograms, preferably from about 25 to about 50 micrograms based uponadministration to a typical 70 kg adult patient.

In vaccine compositions, the AGPs of the subject invention areadministered to a warm-blooded animal, including humans, with anantigen. The amount of antigen administered to elicit a desired responsecan be readily determined by one skilled in the art and will vary withthe type of antigen administered, route of administration andimmunization schedule. For example, 0.2 μg of tetanus toxoidadministered with the claimed compounds subcutaneously to a mouse in twoimmunization 21 days apart elicited a humoral immune response to thatantigen.

The compounds of the subject invention are synthesized by coupling anN-acyloxyacylated or N-protected aminoalkanol or aminoalkanethiol(aglycon unit) with a suitably protected and/or 3-O-acyloxyacylatedglucosaminide unit. In one preferred method for preparing the compoundsof the subject invention (Scheme 1), an N-(2,2,2-trichloroethoxycarbonyl(Troc))-protected glycosyl halide 1 (Z=F, Cl, Br) is coupled with anN-[(R)-3-n-alkanoyloxytetradecanoyl]aminoalkanol or thiol 2 (possessingR₅ and R₆ in suitably protected form) via a Koenigs-Knorr type reactionin the presence of mercury or silver salts to give glycosideintermediate 3. Preferably, the glucosaminide unit 1 possesses ananomeric chloride atom (Z=Cl), and the coupling catalyst is silvertrifluoromethanesulfonate. Intermediate 3 can also be prepared bycoupling the aglycon unit 2 with an N-Troc-protected glycosyl acetate(Z=OAc) or related activated derivative in the presence of a Lewis acidsuch as boron trifluoride etherate. By “activated” is meant having anappropriate displaceable leaving group “Z” attached to the anomericcenter of the glucosaminide unit. Glucosaminide unit 1 bears an(R)-3-n-alkanoyloxytetradecanoyl residue on the 3-position, and suitableprotecting groups on the 6-hydroxyl and 4-phosphate moieties. Typicalprotecting groups for the phosphate group include, but are not limitedto, phenyl, benzyl, and o-xylyl. The phosphate group is protectedpreferably with two phenyl groups. The 6-position can be temporarilyprotected by blocking groups commonly used in sugar chemistry such assilyl, benzyl, or benzyloxymethyl ethers or, alternatively, an alkylcarbonate. The 6-hydroxyl group is protected preferably as a1,1-dimethyl-2,2,2-trichloroethyl carbonate (TCBOC).

The trichloroethyl-based protecting group(s) in the Koenigs-Knorrcoupled product 3 are removed with zinc and the glucosaminide nitrogenis selectively acylated with a (R)-3-n-alkanoyloxytetradecanoic acid 4in the presence of a suitable coupling reagent to give the hexaacylatedderivative 5. The remaining protecting groups in 5 are then cleaved bycatalytic hydrogenation in the presence of a palladium or platinumcatalyst or by other appropriate means to give compounds of Formula (I).

A suitable starting material for the synthesis of glycosyl donor 1 is2-(trimethylsilyl)ethyl2-amino-2-deoxy-4,6-O-isopropylidene-β-D-glucopyranoside which can beprepared from commercially available D-glucosaminide hydrochloride usingpublished procedures. The conversion of the 2-(trimethylsilyl)ethylglycoside starting material to glycosyl donor 1 can be achieved bymethods known in the art or modifications thereof which are describedherein. The aglycon unit 2 can be prepared by N-acyloxyacylation ofcommercially available starting materials with an appropriate(R)-3-n-alkanoyloxytetradecanoic acid 4, or N-acyloxyacylation ofstarting materials that can be obtained by known methods in the chemicalliterature. Alternatively, the N-acyloxyacyl residue in 2 can besubstituted with an appropriate amine protecting group which is removedsubsequent to the coupling reaction such as is described in the secondpreferred embodiment below.

In a second preferred method for preparing the compounds of the subjectinvention (Scheme 2), introduction of the(R)-3-n-alkanoyloxytetradecanoyl and phosphate groups into theglucosaminide and aglycon units is performed subsequent to theglycosylation (coupling) reaction using N- and O-protecting groupssuitable for the chemical differentiation of the amino and hydroxylgroups present. Preferably, the N-Troc-protected glycosyl donor 6 iscoupled with an N-allyloxycarbonyl (AOC)-protected aminoalkanol or thiol7 in the presence of an appropriate catalyst to give the aminoalkylβ-glycoside 8. Most preferably, the glycosyl donor 6 possesses ananomeric acetoxy group (Z=OAc), and the coupling catalyst is borontrifluoride etherate. Other N-protecting groups for the aglycon aminogroup include, but are not limited to, commonly employed carbamatesobvious to one skilled in the art such as t-butyl (t-BOC), benzyl (Cbz),2,2,2-trichloroethyl (Troc), and 9-fluorenylmethyl(Fmoc).

Base-induced cleavage of the acetate groups in coupling product 8 and4,6-acetonide formation under standard conditions known in the art givesintermediate 9. 3-O-Acylation of 9 with (R)-3-n-alkanoyloxytetradecanoicacid 4, followed by palladium(0)-mediated removal of the aglycon N-AOCgroup and N-acylation with (R)-3-n-alkanoyloxytetradecanoic acid 4provides intermediate 10. Acetonide hydrolysis and functionalization ofthe 4- and 6-positions as described herein for the preparation ofglycosyl donor 1 gives intermediate 3 (Y=O) which is then processed asin Scheme 1 to afford compounds of general Formula (I).

The present invention is further described by way of the followingnon-limiting Examples and Test Examples which are given for illustrativepurposes only. It is important to note that the introduction of the(R)-3-n-alkanoyloxytetradecanoyl groups and the phosphate group(s) intothe glucosaminide and aglycon units do not necessarily have to beperformed in the order shown in Schemes 1 and 2 or described in theExamples shown below.

Examples 1-43 describe methods of making the AGP compounds of thesubject invention. Test Examples 1-13 describe assays conducted to thedetermine the immunogenicity of these compounds. Table 1 lists thechemical composition and experimental reference numbers for eachcompound in these examples.

TABLE 1 Example Ref. No. R₁-R₃ n p R₆ q R₇  1 — — — — — — —  2 B1*n-C₁₃H₂₇CO 0 1 OH 0 H  3 B2** n-C₁₃H₂₇CO 0 1 OH 0 H  4 B3 n-C₁₁H₂₃CO 0 1OH 0 H  5 B4 n-C₁₀H₂₁CO 0 1 OH 0 H  6 B5 n-C₉H₁₉CO 0 1 OH 0 H  7 B6***n-C₉H₁₉CO 0 1 OH 0 H  8 B7 n-C₈H₁₇CO 0 1 OH 0 H  9 B8 n-C₆H₁₃CO 0 1 OH 0H 10 B9 n-C₉H₁₉CO 1 1 OH 0 H 11 B10 n-C₉H₁₉CO 0 2 OH 0 H 12 B11n-C₁₃H₂₇CO 0 0 CO₂H 0 H 13 B12 n-C₁₁H₂₃CO 0 0 CO₂H 0 H 14 B13 n-C₁₀H₂₁CO0 0 CO₂H 0 H 15 B14** n-C₉H₁₉CO 0 0 CO₂H 0 H 16 B15* n-C₉H₁₉CO 0 0 CO₂H0 H 17 B16 n-C₈H₁₇CO 0 0 CO₂H 0 H 18 B17 n-C₇H₁₅CO 0 0 CO₂H 0 H 19 B18n-C₆H₁₃CO 0 0 CO₂H 0 H 20 B19 n-C₁₃H₂₇CO 0 0 H 0 H 21 B20 n-C₉H₁₉CO 0 0H 0 H 22 B21 n-C₁₃H₂₇CO 1 0 H 0 H 23 B22 n-C₁₃H₂₇CO 2 0 H 0 H 24 B23n-C₁₃H₂₇CO 4 0 H 0 H 25 B24 n-C₁₃H₂₇CO 0 0 CONH₂ 0 H 26 B25 n-C₉H₁₉CO 00 CONH₂ 0 H 27 B26 n-C₁₃H₂₇CO 0 0 CO₂Me 0 H 28 B27 n-C₁₃H₂₇CO 0 0 H 1CO₂H 29 B28 n-C₉H₁₉CO 1 0 H 1 CO₂H 30 B29 n-C₅H₁₁CO 0 0 CONH₂ 0 H 31 B30R₁ = R₃ = n-C₉H₁₉CO 0 0 CONH₂ 0 H R₂ = n-C₅H₁₁CO 32 B31 n-C₅H₁₁CO 0 0 H0 H 33 B32 R₁ = n-C₁₃H₂₇CO 0 0 H 0 H R₂ = n-C₁₇H₃₅CO R₃ = n-C₁₅H₃₁CO 34B34 n-C₅H₁₁CO 0 0 CO₂H 0 H 35 B35 R₁ = n-C₅H₁₁CO 0 0 CO₂H 0 H R₂ = R₃ =n-C₉H₁₉CO 36 B36 R₁ = R₃ = n-C₉H₁₉CO 0 0 CO₂H 0 H R₂ = n-C₅H₁₁CO 37 B37R₁ = R₂ = n-C₉H₁₉CO 0 0 CO₂H 0 H R₃ = n-C₅H₁₁CO 38 B38 R₁ = n-C₉H₁₁CO 00 CO₂H 0 H R₂ = R₃ = n-C₅H₁₁CO 39 B39 R₁ = R₃ = C₅H₁₁CO 0 0 CO₂H 0 H R₂= n-C₉H₁₉CO 40 B40 R₁ = R₂ = n-C₅H₁₁CO 0 0 CO₂H 0 H R₃ = n-C₉H₁₉CO 41B41 R₁ = R₃ = n-C₉H₁₉CO 0 1 OH 0 H R₂ = n-C₅H₁₁CO 42 B42 n-C₉H₁₁CO 0 2CO₂H 0 H 43 B43 R₁ = n-C₁₃H₂₇CO 0 0 CO₂H 0 H R₂ = n-C₁₁H₂₃CO R₃ = H Forall Examples shown: X = Y = O; R₄ = R₅ = H; m = 0; R₈ = phosphono; R₉ =H. *the stereochemistry of the carbon atom to which R₅ is attached is S.**the stereochemistry of the carbon atom to which R₅ is attached is R.***R₈ is H and R₉ is phosphono.

EXAMPLE 1 PREPARATION OF (R)-3-N-ALKANOYLOXYTETRADECANOIC ACIDS (4).

(1) A solution of methyl 3-oxotetradecanoate (19 g, 0.074 mol) in MeOH(100 mL) was degassed by sparging with argon (15 min).[(R)-Ru(Binap)Cl]₂·NEt₃ catalyst (0.187 g, 0.111 mmol) and 2 N aqueousHCl (0.5 mL) were added and the resulting mixture was hydrogenated at 60psig and 40-50° C. for 18 h. The reaction was diluted with hexanes (250mL), filtered through a short column of silica gel, and concentrated.The crude product was dissolved in tetrahydrofuran (THEF; 200 mL),treated 2.4 N aqueous LiOH (83 mL, 0.2 mol) and stirred vigorously atroom temperature for 4 h. The resulting slurry was partitioned betweenether (200 mL) and 1 N aqueous HCl (200 mL) and the layers separated.The aqueous layer was extracted with ether (100 mL) and the combinedethereal extracts were dried (Na₂SO₄) and concentrated. The crudehydroxy acid was dissolved in hot acetonitrile (250 mL), treated withdicyclohexylamine (DCHA; 17 ml, 0.085 mol) and stirred at 60° C. for 1h. The product that crystallized upon cooling was collected andrecrystallized from acetonitrile (650 mL) to yield 28.6 g (91%) ofdicyclohexylammonium (R)-3-hydroxytetradecanoate as a colorless solid:mp 94-95° C.; ¹H NMR (CDCl₃) δ 0.88 (t, 3 H, J˜6.5 Hz), 1.05-1.58 (m,24H), 1.65 (m, 2H), 1.80 (m, 4H), 2.01 (br d, 4H) 2.18 (dd, 1H, J=15.7,9.4 Hz), 2.36 (dd, 1H, J=15.7, 2.6 Hz), 2.94 (m, 2H), 3.84 (m, 1H).

(2) To a mixture of the compound prepared in (1) above (50 g, 0.117 mol)and 2,4′-dibromoacetophenone (39 g, 0.14 mol) in EtOAc (2.3 L) was addedtriethylamine (19.6 mL, 0.14 mol) and the resulting solution was stirredfor 18 h at room temperature. The voluminous precipitate that formed wascollected and triturated with warm EtOAc (3×400 mL). The combinedtriturates and filtrate were washed with 1 M aq. HCl, saturated aq. NaCland dried (Na₂SO₄). Volatiles were removed under reduced pressure andthe crude product obtained was crystallized from EtOAc-hexanes to give47.2 g (91%) of (R)-3-hydroxytetradecanoic acid p-bromophenacyl ester asa colorless solid: mp 109-109.5° C.; ¹H NMR (CDCl₃) δ 0.88 (˜t, 3H,J˜6.5 Hz) 1.15-1.70 (m, 20H), 2.56 (dd, 1H, J=15.1, 9.1 Hz), 2.69 (dd,1H, J=15.1, 2.9 Hz), 3.27 (br s, 1H), 4.12 (m, 1H), 5.31 (d, 1H, J=16.5Hz), 5.42 (d, 1H, J=16.5 Hz), 7.65 (d, 2H, J=8.5 Hz), 7.78 (d, 2H, J=8.5Hz).

(3) A solution of the compound prepared in (2) above (4.6 g, 10.4 mmol)in CH₂Cl₂ (50 mL) containing 4-dimethylaminopyridine (0.12 g, 1.0 mmol)and pyridine (5 mL, 62 mmol) was treated at room temperature withmyristoyl chloride (3.1 mL, 11.4 mmol). After stirring for 5 h at roomtemperature MeOH (0.5 mL) was added, and the reaction mixture wasconcentrated. The residue was partitioned between Et₂O (150 mL) and cold10% aqueous HCl (50 mL) and the layers separated. The ethereal layer wasdried (Na₂SO₄) and concentrated and the residue obtained was purified ona short pad of silica gel with 5% EtOAc-hexanes. The diester wasdissolved in AcOH (42 mL) and treated with three equal portions of zincdust (6 g, 90 mmol) at 60° C. over a 1 h period. After an additionalhour at 60° C., the cooled reaction mixture was sonicated (5 min),filtered through Celite® and concentrated. The residue was purified byflash chromatography on silica gel with 10% EtOAc-hexanes to give 4.17 g(82%) of (R)-3-tetradecanoyloxytetradecanoic acid as a colorless solid:mp 28-29° C.; ¹H NMR (CDCl₃) δ 0.88 (˜t, 6H), 1.15-1.40 (m, 38H),1.50-1.70 (m, 4H), 2.28 (t, 2H, J=7.4 Hz), 2.56 (dd, 1H, J=15.9, 5.8Hz), 2.63 (dd, 1H, J=15.9, 7.1 Hz), 5.21 (m, 1H).

(4) In the same manner as described in Example 1-(3), the compoundprepared in Example 1-(2) (2.5 g, 5.68 mmol) was acylated with lauroylchloride (1.45 mL, 6.25 mmol) in the presence of pyridine (0.57 mL, 7.0mmol) in CH₂Cl₂ (60 mL) and then deprotected with zinc (9.3 g, 142 mmol)in AcOH (40 mL) to afford (R)-3-dodecanoyloxytetradecanoic acid as acolorless oil: ¹H NMR (CDCl₃) δ 0.90 (t, 6H, J=6.5 Hz), 1.0-1.75 (m,46H), 2.30 (m, 2H), 2.62 (m, 2H), 5.22 (m, 1H).

(5) A solution of the compound prepared in Example 1-(2) (2.5 g, 5.68mmol) was treated with undecanoic acid (1.16 g, 6.25 mmol) and EDCMeI(2.08 g, 7.0 mmol) in CH₂Cl₂ (60 mL) and then deprotected as describedin Example 1-(3) with zinc (9.3 g, 142 mmol) in AcOH (40 mL) to afford(R)-3-undecanoyloxytetradecanoic acid as a colorless oil: ¹H NMR (CDCl₃)δ 0.89 (t, 6H, J=6.7 Hz), 1.0-1.75 (m, 44H), 2.29 (m, 2H), 2.61 (m, 2H),5.22 (m, 1H).

(6) In the same manner as described in Example 1-(3), the compoundprepared in Example 1-(2) (4.4 g, 10 mmol) was acylated with decanoylchloride (2.3 mL, 11 mmol) in the presence of pyridine (1.2 mL, 15.0mmol) in CH₂Cl₂ (100 mL) and then deprotected with zinc (16.4 g, 250mmol) in AcOH (60 mL) to afford (R)-3-decanoyloxytetradecanoic acid as acolorless oil: ¹H NMR (CDCl₃) δ 0.89 (t, 6H, J=6.8 Hz), 1.0-1.75 (m,34H), 2.29 (t, 2H, J=7.4 Hz), 2.61 (t, 2H, J=4.2 Hz), 5.22 (m, 1H).

(7) In the same manner as described in Example 1-(3), the compound 71.prepared in Example 1-(2) (2.5 g, 5.68 mmol) was acylated with nonanoylchloride (1.13 mL, 6.25 mmol) in the presence of pyridine (0.57 mL, 7.0mmol) in CH₂Cl₂ (60 mL) and then deprotected with zinc (9.3 g, 142 mmol)in AcOH (40 mL) to afford (R)-3-nonanoyloxytetradecanoic acid as acolorless oil: ¹H NMR (CDCl₃) δ 0.89 (t, 6H, J=6.9 Hz), 1.0-1.75 (m,32H), 2.29 (t, 2H, J=7.5 Hz), 2.61 (m, 2H), 5.22 (m, 1H).

(8) In the same manner as described in Example 1-(3), the compoundprepared in Example 1-(2) (2.5 g, 5.68 mmol) was acylated with octanoylchloride (1.07 mL, 6.25 mmol) in the presence of pyridine (0.57 mL, 7.0mmol) in CH₂Cl₂ (60 mL) and then deprotected with zinc (9.3 g, 142 mmol)in AcOH (40 mL) to afford (R)-3-octanoyloxytetradecanoic acid as acolorless oil: ¹H NMR (CDCl₃) δ 0.92 (t, 6H, J=6.9 Hz), 1.0-1.75 (m,30H), 2.32 (t, 2H, J=7.4 Hz), 2.63 (t, 2H, J=4.4 Hz), 5.23 (m, 1H).

(9) In the same manner as described in Example 1-(3), the compoundprepared in Example 1-(2) (2.5 g, 5.68 mmol) was acylated with heptanoylchloride (0.97 mL, 6.25 mmol) in the presence of pyridine (0.57 mL, 7.0mmol) in CH₂Cl₂ (60 mL) and then deprotected with zinc (9.3 g, 142 mmol)in AcOH (40 mL) to afford (R)- 3-heptanoyloxytetradecanoic acid as acolorless oil: ¹H NMR (CDCl₃) δ 0.89 (t, 6H, J=6.8 Hz), 1.0-1.75 (m,28H), 2.29 (t, 2H, J=7.4 Hz), 2.61 (d, 2H, J=5.8 Hz), 5.22 (m, 1H).

EXAMPLE 2 B1 PREPARATION OF3-HYDROXY-(S)-2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]PROPYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMIO]-3-O-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₃H₂₇CO, X═Y═O,N=M=Q=0, R₄═R₅═R₇═R₉═H, R₆═OH, P=1, R₈═PO₃H₂).

(1) To a solution of 2-(trimethylsilyl)ethyl2-amino-2-deoxy-4,6-O-isopropylidene-β-D-glucopyranoside (6.46 g, 20.2mmol) in CHCl₃ (300 mL) was added 1 N aqueous NaHCO₃ (300 mL) and2,2,2-trichloroethyl chloroformate (8.5 g, 40 mmol). The resultingmixture was stirred vigorously for 3 h at room temperature. The organiclayer was separated, dried (Na₂SO₄) and concentrated to give a colorlesssyrup. Flash chromatography on silica gel (gradient elution, 30→40%EtOAc-hexanes) afforded 9.6 g (96%) of 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless solid: mp 69-70° C.; ¹H NMR (CDCl₃) δ 0.0 (s, 9H), 0.94(m, 2H), 1.44 and 1.52 (2s, 6H), 2.94 (br s, 1H), 3.23-3.37 (m, 2H),3.48-3.62 (m, 2H), 3.79 (t, 1H, J=10.5 Hz), 3.88-4.08 (m, 3H), 4.65 (d,1H, J=8.3 Hz), 4.74 (m, 2H), 5.39 (d, 1H, J=7.4 Hz).

(2) A solution of the compound prepared in (1) above (7.5 g, 15.2 mmol),(R)-3-tetradecanoyloxytetradecanoic acid (7.58 g, 16.7 mmol) and4-pyrrolidinopyridine (0.25 g, 1.7 mmol) in CH₂Cl₂ (95 mL) was treatedwith 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide methiodide (EDC·MeI;4.94 g, 16.7 mmol) and stirred for 16 h at room temperature. Thereaction mixture was filtered through a short pad of Celite®,concentrated, and the resulting residue was heated at 60° C. in 90%aqueous AcOH (100 mL) for 1 h. The mixture was concentrated and residualAcOH and water were removed by azeotroping with toluene (2×150 mL). Thecrude diol was purified by flash chromatography on silica gel (gradientelution, 30→40% EtOAc-hexanes) to give 11.8 g (83%) of2-(trimethylsilyl)ethyl2-deoxy-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.0 (s, 9H), 0.9 (m, 8H),1.1-1.7 (m, 42H), 2.30 (t, 2H, J=7.4 Hz), 2.52 (m, 2H), 3.36-3.72 (m,4H), 3.78-4.03 (m, 3H), 4.57 (d, 1H, J=8.3 Hz), 4.65 (d, 1H, J=11 Hz),4.77 (d, 1H, J=11 Hz), 5.0-5.15 (m, 2H), 5.20 (d, 1H, J=7.4 Hz).

(3) A solution of the compound prepared in (2) above (10.9 g, 12 mmol)and pyridine (2 mL, 25 mmol) in CH₂Cl₂ (125 mL) at 0° C. was treateddropwise over 15 min with a solution of2,2,2-trichloro-1,1-dimethylethyl chloroformate (3.17 g, 13.2 mmol) inCH₂Cl₂ (25 mL). The reaction mixture was allowed to warm slowly toambient temperature over 3.5 h. 4-Pyrrolidinopyridine (0.89 g, 6.0mmol), N,N-diisopropylethylamine (10.5 mL, 60 mmol) and diphenylchlorophosphate (3.7 mL, 18 mmol) were added sequentially and theresulting mixture was stirred for 5 h at room temperature. The reactionmixture was diluted with CH₂Cl₂ (500 mL), washed with cold 7.5% aqueousHCl (2×250 mL), water (250 mL), saturated aqueous NaHCO₃ (250 mL), dried(Na₂SO₄), and then concentrated. The residue obtained was purified byflash chromatography on silica gel eluting with 12.5% EtOAc-hexanes togive 15.1 g (95%) of 2-(trimethylsilyl)ethyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichlorethoxycarbonylamnino)-β-D-glucopyranosideas a viscous oil: ¹H NMR (CDCl₃) δ 0.0 (s, 9H), 0.8-1.0 (m, 8H),1.1-1.65 (m, 42H), 1.83 and 1.90 (2s, 6H), 2.15-2.45 (m, 4H), 3.34 (q,1H, J=8 Hz), 3.37 (m, 1H), 3.81 (m, 1H), 3.95 (m, 1 H), 4.27 (dd, 1H,J=12, 5 Hz), 4.34 (d, 1H, J=12 Hz), 4.58 (d, 1H, J=12 Hz), 4.66 (q, 1H,J=9 Hz), 4.86 (d, 1H, J=12 Hz), 5.03 (d, 1H, J=7.9 Hz), 5.21 (m, 1H),5.54-5.70 (m, 2H), 7.2-7.8 (mn, 10H).

(4) A solution of the compound prepared in (3) above (1.87 g, 1.41 mmol)in CH₂Cl₂ (3 mL) at 0° C. was treated dropwise over 10 min withtrifluoroacetic acid (TFA; 6 mL) and then stirred for 4 h at 0° C. Thereaction mixture was concentrated and residual TFA was removed byazeotroping with toluene (2×5 mL). A solution of the lactol anddimethylformamide (2.2 mL, 28.2 mmol) in CH₂Cl₂ (14 mL) at 0° C. wastreated with oxalyl bromide (2.0 M in CH₂Cl₂; 2.1 mL, 4.2 mmol) dropwiseover 15 min and the resulting suspension was stirred at 0° C. for 24 h.The reaction mixture was partitioned between cold saturated aqueousNaHCO₃ (25 mL) and ether (50 mL) and the layers were separated. Theethereal layer was washed with saturated aqueous NaCl, dried (Na₂SO₄)and concentrated to give 1.85 g (˜100%) of2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosylbromide as a colorless glass.

(5) A solution of (R)-2-amino-3-benzyloxy-1-propanol (0.46 g, 2.33 mmol)and (R)-3-tetradecanoyloxytetradecanoic acid (1.29 g, 2.83 mmol) inCH₂Cl₂ (20 mL) was treated with EDC·MeI (0.78 g, 2.79 mmol) and stirredfor 16 h at room temperature. The reaction mixture was filtered througha short pad of Celite® and concentrated. Flash chromatography on silicagel with 45% EtOAc-hexanes afforded 1.1 g (69%) of3-benzyloxy-(R)-2-[(R)-3-tetradecanoyloxytetradecanoylamino]propanol asa colorless solid: mp 42-44.5° C.; ¹H NMR δ 0.88 (t, 6H, J=6.5 Hz),1.0-1.7 (m, 42 H), 2.50 (t, 2H, J=7.5 Hz), 2.46 (m, 2H), 3.56 (br s,1H), 3.5-3.75 (m, 3H), 3.78 (dd, 1H, J=11, 4 Hz), 4.08 (m, 1H), 4.51 (s,2H), 5.17 (m, 1H), 6.36 (d, 1H, J=7.8 Hz), 7.2-7.4 (m, 5H).

(6) To a solution of the compound prepared in (4) above (1.00 g, 0.776mmol) and the compound prepared in (5) above (0.35 g, 0.57 mmol) indichloroethane (4.5 mL) was added powdered 4 A molecular sieves (1.25 g)and calcium sulfate (2.7 g, 20 mmol). After stirring for 10 min at roomtemperature, the mixture was treated with mercury cyanide (1.0 g, 4.0mmol) and then heated to reflux for 12 h shielded from light. Thereaction mixture was diluted with CH₂Cl₂ (25 mL) and filtered through apad of Celite®. The filtrate was washed with 1N aqueous KI (25 mL),dried (Na₂SO₄) and concentrated. The residue was chromatographed onsilica gel with EtOAc-hexanes-MeOH (80:20:0→70:30:1, gradient elution)to give 0.66 g (63%) of3-benzyloxy-(S)-2-[(R)-3-tetradecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-phosphono-3-O-[(R)-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR δ 0.88 (t, 12H, J=6.5 Hz), 1.0-1.65 (m,84H), 1.79 and 1.86 (2s, 6H), 2.1-2.5 (m, 8H), 3.35-3.55 (m, 3H),3.65-3.8 (mn, 3H), 4.1-4.75 (m, 9H), 5.05-5.3 (m, 2H), 5.3-5.5 (m, 2H),6.04 (d, 1H, J=8.4 Hz), 7.05-7.45 (m, 15H).

(7) A stirred solution of the compound prepared in (6) above (0.60 g,0.328 mmol) in AcOH (9 mL) at 55° C. was treated with zinc dust (1.1 g,16 mmol) in three equal portions over 1 h. The cooled reaction mixturewas sonicated, filtered through a bed of Celite® and concentrated. Theresulting residue was partitioned between CH₂Cl₂ (60 mL) and cold 1 Naqueous HCl (35 mL) and the layers separated. The organic layer waswashed with 5% aqueous NaHCO₃, dried (Na₂SO₄) and concentrated. Amixture of the residue obtained and (R)-3-tetradecanoyloxytetradecanoicacid (0.18 g, 0.39 mmol) in CH₂Cl₂ (3.5 mL) was stirred with powdered 4Å molecular sieves (0.1 g) for 30 min at room temperature and thentreated with 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ; 0.12g, 0.49 mmol). The resulting mixture was stirred for 6 h at roomtemperature, filtered through Celite® and then concentrated.Chromatography on silica gel (gradient elution, 0.5→1% MeOH—CHCl₃)afforded 0.31 g (50%) of3-benzyloxy-(S)-2-[(R)-3-tetradecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-2-[(R)-3-tetradecanoyloxytetradecanoylamino]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 18H, J=˜6.5 Hz),1.0-1.8 (m, 126H), 2.1-2.5 (m, 12H), 3.35-3.75 (m, 6H), 3.80 (m, 2H),4.23 (m, 1H), 4.46 (d, 1H, J=12 Hz), 4.51 (d, 1H, J=12 Hz), 4.65 (q, 1H,J=˜9.5 Hz), 4.82 (d, 1H, J=8.1 Hz), 5.05-5.25 (m, 3H), 5.47 (t, 1H,J=˜9.5 Hz), 6.16 (d, 1H, J=8.1 Hz), 6.31 (d, 1H, J=8.4 Hz), 7.1-7.4 (m,15H).

(8) A solution of the compound prepared in (7) above (0.26 g, 0.138mmol) in THF (25 mL) was hydrogenated in the presence of 5% palladium oncarbon (50 mg) at room temperature and atmospheric pressure for 16 h.After removal of the catalyst by filtration, AcOH (3 mL) and platinumoxide (0.14 g) were added and the hydrogenation was continued at roomtemperature and 75 psig for 24 h. The resulting opalescent reactionmixture was diluted with 2:1 CHCl₃—MeOH (20 mL) and sonicated briefly togive a clear solution. The catalyst was collected, washed with 2:1CHCl₃-MeOH (2×5 mL) and the combined filtrate and washings wereconcentrated. The residue was dissolved in 1% aqueous triethylamine (10mL) by sonicating for 5 min at 35° C. and the resulting solution waslyophilized. Flash chromatography on silica gel withchloroform-methanol-water-triethylamine (94:6:0.5:0.5→88:12:1.0:1.0,gradient elution) afforded 0.20 g (84%) of product as a colorlesspowder. A portion of the chromatography product (0.166 g) was dissolvedin cold 2:1 CHCl₃—MeOH (33 mL) and washed with cold 0.1 N aqueous HCl(14 mL). The lower organic layer was filtered and concentrated and thefree acid obtained was lyophilized from 1% aqueous triethylamine(pyrogen free, 15 mL) to give 0.160 g of3-hydroxy-(S)-2-[(R)-tetradecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-phosphono-2-[(R)-3-tetradecanoyloxytetradecanoylamino]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as a colorless solid: mp 178-180° C. (dec); IR(film) 3293, 3103, 2959, 2924, 2855, 1732, 1654, 1640, 1553, 1467, 1377,1259, 1175, 1106, 1086, 1050, 803, 720 cm⁻¹; HMR (CDCl₃—CD₃OD) δ 0.88(t, 18H, J=˜7 Hz), 1.0-1.7 (mn, 135H), 2.15-2.75 (m, 12H), 3.02 (q, 6H,J=7 Hz), 3.35-4.1 (m, 7H), 4.22 (q, 1H, J=˜9.5 Hz), 4.77 (d, 1H, J=8Hz), 5.05-5.35 (mn, 4H), 6.58 (d, 1H, J=6 Hz), 6.73 (d, 1H, J=7.5 Hz,NH); ¹³C NMR (CDCl₃) δ 173.5, 173.2, 170.7, 170.5, 170.0, 100.7, 75.9,72.7, 71.2, 71.0, 70.8, 70.6, 67.9, 61.7, 60.5, 55.0, 50.4, 45.6, 41.4,39.5, 34.5, 34.4, 32.0, 31.8, 30.3, 29.8, 29.4, 29.3, 25.3, 25.1, 22.7,14.2, 8.6.

Anal. Calcd for C₁₉H₁₉₂N₃O₁₈P.5H₂O: C, 64.84;H, 11.10; N, 2.29; P, 1.69.Found: C, 64.69;H, 11.24; N, 1.93; P, 1.44.

EXAMPLE 3 B2 PREPARATION OF3-HYDROXY-(R)-2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]PROPYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₃H₂₇CO, X═Y═O,N=M=Q=0, R₄═R₅═R₇═R₉═H, R₆═OH, P=1, R₈═PO₃H₂).

(1) A solution of the compound prepared in Example 2-(5) (0.63 g, 1.02mmol) in CH₂Cl₂ (7 mL) was treated sequentially with pyridine (0.4 mL, 5mmol), 4-dimethylaminopyridine (cat.) and2,2,2-trichloro-1,1-dimethylethyl chloroformate (0.307 g, 1.23 mmol) andstirred for 16 h at room temperature. The reaction mixture was dilutedwith CH₂Cl₂ (25 mL), washed with saturated aqueous NaHCO₃ (25 mL) anddried (Na₂SO₄). Removal of volatiles in vacuo gave a residue which wasdissolved in THF-AcOH (10 mL, 9:1) and hydrogenated in the presence of5% palladium on carbon (150 mg) at room temperature and atmosphericpressure for 24 h. After removal of the catalyst by filtration andconcentration of the filtrate, the residue was purified by flashchromatography on silica gel with 35% EtOAc-hexanes to give 0.536 g(72%) of3-(2,2,2-trichloro-1,1-dimethylethoxycarbonyloxy)-(S)-2-[(R)-3-tetradecanoyloxytetradecanoylamino]propanolas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=˜6.5 Hz), 1.1-1.7(m, 42H), 1.94 (s, 6H), 2.30 (t, 2H, J=7.5 Hz), 2.47 (d, 2H, J=6 Hz),3.50 (br s, 1H), 3.72 (m, 2H), 4.15-4.35 (m, 3H), 5.15 (m, 1H), 6.18 (d,1H, J=7.2 Hz).

(2) In the same manner as described in Example 2-(6), the compoundprepared in (1) above (0.310 g, 0.426 mmol) and the compound prepared inExample 2-(4) (0.961 g, 0.745 mmol) were coupled in the presence ofmercury cyanide (0.43 g, 1.7 mmol) to give 0.644 g (78%) of3-(2,2,2-trichloro-1,1-dimethylethyloxycarbonyloxy)-(S)-2-[(R)-3-tetradecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-phosphono-3-O-[(R)-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=˜6.5 Hz),1.0-1.7 (m, 84H), 1.81 and 1.89 (2s, 6 H), 1.93 (s, 6H), 2.15-2.55 (m,8H), 3.45-3.7 (m, 2H), 3.80 (br d, 1H, J=9 Hz), 3.9-4.45 (m, 6H),4.6-4.8 (m, 3H), 4.87 (d, 1H, J=8.1 Hz), 5.0-5.25 (m, 2H), 5.48 (t, 1H,J=˜9.5 Hz), 6.1-6.3 (m, 2H).

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (0.602 g, 0.310 mmol) was deprotected with zinc(1.5 g, 23 mmol) and acylated with (R)-3-tetradecanoyloxytetradecanoicacid, (0.17 g, 0.37 mmol) in the presence of EEDQ (0.115 g, 0.467 mmol)to give 0.365 g (66%) of3-hydroxy-(R)-2-[(R)-3-tetradecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-2-[(R)-3-tetradecanoyloxytetradecanoylamino]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 18H, J=˜6.5 Hz),1.0-1.7 (m, 126H), 2.15-2.55 (m, 12H), 3.18 (br s, 1H), 3.45-3.8 (m,8H), 3.85-4.05 (m, 2H), 4.69 (q, 1H, J=9.5 Hz), 5.05-5.25 (m, 3H), 5.42(t, 1H, J=˜9.5 Hz), 6.42 (d, 1H, J=7.8 Hz), 6.59 (d, 1H, J=7.2 Hz),7.1-7.4 (m, 10H).

(4) In the same manner as described in Example 2-(8), the compoundprepared in (3) above (0.355 g, 0.196 mmol) was hydrogenated in thepresence of platinum oxide (175 mg) to give 0.265 g (77%) of3-hydroxy-(R)-2-[(R)-3-tetradecanoyloxytetradecanoylamino]propyl2deoxy-4-O-phosphono-2-[(R)-3-tetradecanoyloxytetradecanoylamino]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as a colorless solid: mp 159-160° C.; IR (film)3291, 2956, 2922, 2853, 1738, 1732, 1716, 1650, 1643, 1556, 1468, 1171,1109, 1083, 1051 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=˜6.5 Hz),1.0-1.7 (m, 135H), 2.15-2.75 (m, 12H), 3.06 (q, 6H, J=7 Hz), 3.25-3.45(m, 2H), 3.5-4.05 (m, 12H), 4.19 (q, 1H, J=˜9.5 Hz), 4.48 (d, 1H, J=8.4Hz), 5.04-5.26 (m, 4H), 7.18 (d, 1H, J=7.8 Hz), 7.27 (d, 1H, J=8.7 Hz);¹³C NMR (CDCl₃) δ 173.5, 173.4, 170.7, 170.6, 170.1, 101.0, 76.0, 72.6,71.4, 71.0, 70.8, 70.6, 68.7, 61.8, 60.5, 55.3, 50.5, 45.6, 41.5, 41.4,39.5, 34.6, 34.4, 34.3, 32.0, 29.8, 29.4, 25.4, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd for C₉₉H₁₉₂N₃O₁₈P.H₂O: C, 67.50;H, 11.10; N, 2.39; P, 1.76.Found: C, 67.40;H, 11.22; N, 2.34; P, 2.11.

EXAMPLE 4 B3 PREPARATION OF3-HYDROXY-(S)-2-[(R)-3-DODECANOYLOXYTETRADECANOYLAMINO]PROPYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DODECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DODECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₁H₂₃CO, X═Y═O,N=M=Q=0, R₄═R₅═R₇═R₉═H, R₆═OH, P=1, R₈═PO₃H₂).

(1) A solution of D-glucosaminide hydrochloride (20 g, 92.8 mmol) in H₂O(250 mL) was treated with a saturated aqueous NaHCO₃ (250 mL) and2,2,2-trichloroethyl chloroformate (14.05 mL, 102 mmol) and stirredvigorously for 18 h. The white solid that formed was collected on afritted funnel and dried under vacuum for 24 h. A solution of the solidin pyridine (100 mL) was cooled to 0° C. and treated with aceticanhydride (100 mL) via addition funnel. The solution was stirred for 18h at room temperature, poured into 1 L of H₂O and extracted with CHCl₃(3×500 mL). The solvent was removed in vacuo to afford 45 g (quant.) ofN-(2,2,2-trichloroethoxycarbonylamino)-1,3,4,6-tetra-O-acetyl-2-deoxy-α-D-glucopyranosidewhich was used without further purification: ¹H NMR (CDCl₃) δ 2.06 (s,6H), 2.12 (s, 3H), 2.22 (s, 3H), 4.03 (m, 1H), 4.07 (d, 1H, J=12.4 Hz),4.22 (dt, 1H, J=9.9, 3.6 Hz), 4.30 (dd, 1H, J=12.4, 4.0 Hz), 4.64 (d,1H, J=9.6 Hz), 5.28 (dt, 1H, J=10.2, 9.9 Hz), 6.25 (d, 1H, J=3.6 Hz).

(2) A solution of (R)-2-amino-3-benzyloxy-1-propanol (5 g, 27.6 mmol) inCH₂Cl₂ (250 mL) was treated with allyl chloroformate (3.2 mL, 30 mmol)and saturated aqueous NaHCO₃ (250 mL) for 18 h. The organic layer wasseparated and concentrated in vacuo. Purification by chromatographyeluting with 30% EtOAc/hexanes afforded 6.9 g (94%) of(R)-2-(allyloxycarbonylamino)-3-benzyloxy-1-propanol as an amorphoussolid: ¹H NMR (CDCl₃) δ 2.56 (br s, 1H), 3.69 (m, 3H), 3 88 (m, 2H),4.54 (s, 2H), 4.58 (d, 2H, J=5.6 Hz), 5.23 (dd, 1H, J=10.4, 1.1 Hz),5.33 (dd, 1H, J=17.1, 1.1 Hz), 5.42 (m, 1H), 5.93 (m, 1H), 7.35 (m, 5H).

(3) A solution of the compounds prepared in (1) and (2) above (8.9 g, 17mmol and 3.6 g, 10 mmol, respectively) in CH₂Cl₂ was treated with borontrifluoride etherate (4.3 mL, 34 mmol) at room temperature for 16 h. Thereaction mixture was quenched with saturated aq. NaHCO₃ (100 mL) andextracted with EtOAc (3×100 mL). The combined EtOAc extracts were dried(Na₂SO₄) and concentrated. The residue obtained was chromatographed with20% EtOAc/hexanes to afford 6.03 g (83%) of3-benzyloxy-(S)-2-(allyloxycarbonylarnino)propyl2-deoxy-3,4,6-tri-O-acetyl-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 2.02 (s, 3H), 2.03 (s, 3H), 2.08(s, 3H), 3.45 (m, 1H), 3.54 (m, 1H), 3.64 (m, 1H), 3.76 (d, 1H, J=7.2Hz), 3.91 (m, 2H), 4.12 (d, 1H, J=12.2 Hz), 4.26 (dd, 1H, J=12.4, 4.7Hz), 4.37 (d, 1H, J=8.2 Hz), 4.43 (d, 1H, J=12.1 Hz), 4.55 (m, 2H), 4.68(m, 2H), 4.87 (d, 1H, J=8.0 Hz), 5.07 (m, 2H), 5.21 (d, 1H, J=9.7 Hz),5.29 (d, 1H, J=17.3 Hz), 5.91 (m, 1H), 7.36 (m, 5H).

(4) A solution of the compound prepared in (3) above (6.0 g, 8.3 mmol)in methanol (83 mL) was treated with ammonium hydroxide (8.3 mL) at roomtemperature for 2 h. The solvent was removed in vacuo and replaced with2,2-dimethoxypropane (50 mL) and camphorsulfonic acid (100 mg) wasadded. The reaction was stirred for 18 h, neutralized with solid NaHCO₃(1 g), filtered and concentrated in vacuo. Purification bychromatography with 50% EtOAc/hexanes afforded 4.58 g (86%) of3-benzyloxy-(S)-2-(allyloxycarbonylamino)propyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranoside:¹H NMR (CDCl₃) δ 1.46 (s, 3H), 1.53 (s, 3H), 2.94 (m, 1H), 3.25 (m, 1H),3.55 (m, 4 H), 3.83 (m, 3H), 3.93 (m, 3H), 4.52 (m, 5H), 4.68 (d, 1H,J=12.1 Hz), 4.77 (d, 1H, J=12.1 Hz), 5.07 (m, 1H), 5.26 (m, 2H), 5.92(m, 1H), 7.37 (m, 5H).

(5) A solution of the compound prepared in (4) above (1.0 g, 1.56 mmol)in CH₂Cl₂ (20 mL) was treated with (R)-3-dodecanoyloxytetradecanoic acid(730 mg, 1.71 mmol) in the presence of EDC·MeI (560 mg, 1.87 mmol) and4-pyrrolidinopyridine (50 mg). The reaction was stirred at roomtemperature for 18 h and filtered through a 6×8 cm plug of silica gelusing 20% EtOAc/hexanes as eluent to afford 1.33 g (82%) of3-benzyloxy-(S)-2-(allyloxycarbonylamino)propyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-dodecanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.8 Hz), 1.1-1.6(m, 38H), 1.37 (s, 3H), 1.46 (s, 3H), 2.28 (t, 2H, J=7.4 Hz), 2.49 (dd,1H, J=15.1, 6.0 Hz), 2.61 (dd, 1H, J=15.1, 6.6 Hz), 3.25-4.0 (m, 9H),4.38 (m, 2H), 4.54 (m, 2H), 4.65 (m, 2H), 4.97 (m, 2H), 5.25 (m, 5H),5.88 (m, 1H), 7.34 (m, 5H).

(6) To a solution of the compound prepared in (5) above (1.31 g, 1.25mmol) in THF (20 mL) was added dimethyl malonate (1.0 mL, 0.88 mmol) andthe solution was degassed in a stream of argon for 30 min.Tetrakis(triphenylphosphine)palladium(0) (200 mg) was added and thereaction was stirred at room temperature for 2 h, and then concentratedin vacuo. The residue obtained was chromatographed on silica gel elutingwith 5-10% EtOAc/CHCl₃. The free amine obtained was acylated with(R)-3-dodecanoyloxytetradecanoic acid (560 mg, 1.38 mmol) in thepresence of EEDQ (370 mg, 1.5 mmol) in CH₂Cl₂ (15 mL). After stirring atroom temperature for 18 h, the solvent was removed in vacuo and theresultant oil was chromatographed on silica gel eluting with20%EtOAc/hexanes to afford 1.02 g (63%) of3-benzyloxy-(S)-2-[(R)-3-dodecanoyloxytetradecanoylamino]propyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-dodecanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.9Hz), 1.1-1.7 (m, 78H), 1.38 (s, 3H), 1.46 (s, 3H), 2.26 (m, 4H), 2.49(dd, 1H, J=15.1, 6.0 Hz), 2.61 (dd, 1H, J=15.1, 6.6 Hz), 3.25-4.0 (m,9H), 5.01 (m, 2H), 6.02 (d, 1H, J=8.4 Hz), 7.34 (m, 5H).

(7) The compound prepared in (6) above (1.0 g, 0.78 mmol) was treatedwith 90% aqueous AcOH (20 mL) for 1 h at 60° C. The solution wasconcentrated in vacuo and residual AcOH and H₂O were removed byazeotroping with toluene (10 mL). The residue was dissolved in CH₂Cl₂,cooled to 0° C., and treated with pyridine (0.076 mL, 0.94 mmol) and asolution of 2,2,2-trichloro-1,1-dimethylethyl chloroformate (205 mg,0.86 mmol) in CH₂Cl₂ (5 mL). The reaction mixture was then allowed towarm and stir at room temperature for 18 h. The resulting light yellowsolution was treated with diphenyl chlorophosphate (0.24 mL, 1.17 mmol),triethylamine (0.22 mL, 1.56 mmol) and catalytic 4-pyrrolidinopyridine(50 mg), and then stirred an additional 24 h at room temperature. Thereaction mixture was diluted with Et₂O (100 mL) and washed with 10% aq.HCl (50 mL). The organic phase was separated, dried over Na₂SO₄ andconcentrated in vacuo. Chromatography over silica gel using 10%EtOAc/hexanes afforded 1.13 g (85%) of3-benzyloxy-(S)-2-[(R)-3-dodecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-dodecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.87 (t, 12H, J=6.9Hz), 1.1-1.6 (m, 78H), 1.78 (s, 3H), 1.86 (s, 3H), 2.01 (m, 1H), 2.18(m, 3H), 2.40 (m, 2H), 2.67 (m, 1H), 2.88 (d, 1H, J=6.6 Hz), 2.97 (d,1H, J=6.9 Hz), 3.41 (m, 2H), 3.72 (m, 1H), 3.82 (m, 1H), 4.24 (m, 1H),4.42 (d, 1H, J=11.8 Hz), 4.64 (m, 3H), 5.16 (m, 1H), 5.39 (m, 2H), 5.75(d, 1H, J=4.3 Hz), 6.05 (d, 1H, J=8.4 Hz), 7.23 (m, 15H).

(8) In the same manner as described in Example 2-(7), the compoundprepared in (7) above (1.1 g, 0.65 mmol) was deprotected with zinc (2.1g, 32 mmol) and acylated with (R)-3-dodecanoyloxytetradecanoic acid (330mg, 0.78 mmol) in the presence of EEDQ (230 mg, 0.94 mmol) to afford 399mg (37%) of3-benzyloxy-(S)-2-[(R)-3-dodecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-2-[(R)-3-dodecanoyloxytetradecanoylamino]-3-O-[(R)-3-dodecanoyltetradecanoyl]-β-D-glucopyranosideas a colorless amorphous solid.

(9) In the same manner as described in Example 2-(8), the compoundprepared in (8) above (399 mg, 0.24 mmol) was hydrogenated in thepresence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) andplatinum oxide (300 mg) in EtOH/AcOH (10:1) to afford 65 mg (16%) of3-hydroxy-(S)-2-[(R)-3-dodecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-phosphono-2-[(R)-3-dodecanoyloxytetradecanoylamino]-3-O-[(R)-3-dodecanoyloxytetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as a white powder: mp 181-184° C. (dec): IR (film)3306, 2956, 2922, 2852, 1732, 1644, 1549, 1467, 1377, 1164, 1106, 1051,721 cm⁻¹; ¹HNMR(CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.7 Hz), 1.1-1.7(m,123H), 2.2-2.7 (m, 12H), 3.06 (q, 6H, J=7.1 Hz), 3.3-4.0 (m, 13H), 4.23(m, 1H), 4.44 (d, 1 H, J=7.7 Hz), 5.0-5.3 (m, 4H); ¹³C NMR (CDCl₃) δ173.9, 173.5, 173.3, 170.8, 170.5, 170.1, 101.0, 75.5, 73.0, 71.1, 70.9,70.6, 67.9, 61.6, 60.7, 54.4, 50.4, 45.8, 41.6, 41.4, 39.6, 34.6, 31.9,29.7, 29.4, 29.3, 25.4, 25.1, 22.7, 14.2, 8.6.

Anal. Calcd. for C₉₃H₁₅₀N₃O₁₈P.H₂O: C, 66.59;H, 10.94; N, 2.50; P, 1.85.Found: C, 66.79;H, 10.65; N, 2.36; P, 1.70.

EXAMPLE 5 B4 PREPARATION OF3-HYDROXY-(S)-2-[(R)-3-UNDECANOYLOXYTETRADECANOYLAMINO]PROPYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-UNDECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-UNDECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₀H₂₁CO, X═Y═O,N=M=Q=0, R₄═R₅═R₇═R₉═H, R₆═OH, P=1, R₈═PO₃H₂).

(1) In the same manner as described in Example 4-(5), the compoundprepared in Example 4-(4) (1.0 g, 1.56 mmol) was acylated with(R)-3-undecanoyloxytetradecanoic acid (705 mg, 1.71 mmol) in thepresence of EDC·MeI (560 mg, 1.87 mmol) and 4-pyrrolidinopyridine (50mg) in CH₂Cl₂ (20 mL) to afford 1.23 g (77%) of3-benzyloxy-(S)-2-(allyloxycarbonylamino)propyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-undecanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, =6.9 Hz), 1.1-1.6(m, 36H), 1.37 (s, 3H), 1.46 (s, 3H), 2.28 (m, 2H), 2.52 (dd, 1H,J=15.1, 6.0 Hz), 2.61 (dd, 1H, =15.5, 6.8 Hz), 3.25 (m, 1H), 3.35-4.0(m, 9H), 4.31 (m, 2H), 4.54 (m, 2H), 4.64 (m, 2H), 5.02 (m, 2H), 5.18(m, 2H), 5.25 (m, 1H), 5.86 (m, 1H), 7.34 (m, 5H). (2) In the samemanner as described in Example 4-(6) the compound prepared in (1) above(1.21 g, 1.17 mmol) was deprotected in THF (20 mL) in the presence ofdimethyl malonate (1.0 mL, 0.88 mmol) andtetrakis(triphenylphosphine)palladium(0) (200 mg) and then acylated with(R)-3-undecanoyloxytetradecanoic acid (540 mg, 1.30 mmol) in thepresence of EEDQ (370 mg, 1.5 mmol) to afford 921 mg (61%) of3-benzyloxy-(S)-2-[(R)-3-undecanoyloxytetradecanoylamino]propyl2-deoxy4,6-O-isopropylidene-3-O-[(R)-3-undecanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.6Hz), 1.1-1.7 (m, 72H), 1.38 (s, 3H), 1.46 (s, 3H), 2.26 (m, 3H), 2.38(m, 5H), 2.49 (dd, 1H, J=15.2, 6.0 Hz), 2.61 (dd, 1H, J=15.0, 6.5 Hz),3.25-4.0 (m, 9H), 4.30 (m, 2H), 4.59 (m, 3H), 6.03 (d, 1H, J=8.2 Hz),7.34 (m, 5H).

(3) In the same manner as described in Example 4-(7) the compoundprepared in (2) above (910 g, 0.71 mmol) was deprotected in 90% aqueousAcOH (20 mL), and then treated with pyridine (0.071 mL, 0.88 mmol) and2,2,2-trichloro-1,1-dimethylethyl chlorofonnate (195 mg, 0.80 mmol) inCH₂Cl₂ followed by diphenyl chlorophosphate (0.23 mL, 1.10 mmol),triethylamine (0.20 mL, 1.46 mmol) and catalytic 4-pyrrolidinopyridine(50 mg) to afford 1.10 g (89%) of3-benzyloxy-(S)-2-[(R)-3-undecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-undecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: H NMR (CDCl₃) δ 0.87 (t, 12H, J=6.7 Hz),1.1-1.6 (m, 72H), 1.78 (s, 3H), 1.86 (s, 3H), 2.01 (m, 1H), 2.18 (m,3H), 2.40 (m, 2H), 2.67 (m, 1H), 2.88 (d, 1H, J=6.7 Hz), 2.97 (d, 1H,J=6.9 Hz), 3.41 (m, 2H), 3.72 (m, 1H), 3.82 (m, 1H), 4.24 (m, 1H), 4.42(d, 1H, J=11.8 Hz), 4.64 (m, 3H), 5.16 (m, 1 H), 5.39 (m, 2H), 5.75 (d,1H, J=4.6 Hz), 6.05 (d, 1H, J=8.4 Hz), 7.22 (m, 15H).

(4) In the same manner as described in Example 2-(7), the compoundprepared in (3) above (1.0 g, 0.59 mmol) was deprotected with zinc (2.0g, 30 mmol) and acylated with (R)-3-undecanoyloxytetradecanoic acid (292mg, 0.71 mmol) in the presence of EEDQ (210 mg, 0.85 mmol) to afford 388mg (40%) of3-benzyloxy-(S)-2-[(R)-3-undecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-2-[(R)-3-undecanoyloxytetradecanoylamino]-3-O-[(R)-3-undecanoyltetradecanoyl]-β-D-glucopyranosideas a colorless amorphous solid.

(5) In the same manner as described in Example 2-(8), the compoundprepared in (4) above (388 mg, 0.24 mmol) was hydrogenated in thepresence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) andplatinum oxide (300 mg) in EtOH/AcOH (10:1) to afford 65 mg (17%) of3-hydroxy-(S)-2-[(R)-3-undecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-phosphono-2-[(R)-3-undecanoyloxytetradecanoylamino]-3-O-[(R)-3-undecanoyloxytetradecanoyl]-β-D-glucopyranosidetnrethylamnonium salt as a white powder: mp 183-184° C; IR (film) 3306,2956, 2922, 2852, 1732, 1644, 1550, 1467, 1377, 1164, 1106, 1052, 721cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.8 Hz), 1.1-1.7 (m, 117H),2.2-2.7 (m, 12H), 3.07 (q, 6H, J=7.1 Hz), 3.3-3.9 (m, 13H), 4.23 (m,1H), 4.45 (d, 1H, J=8.2 Hz), 5.0-5.3 (m, 4H); ¹³C NMR (CDCl₃) δ 173.8,173.5, 173.3, 170.8, 170.5, 170.1, 101.0, 75.5, 73.1, 71.5, 71.3, 70.9,70.6, 67.8, 61.6, 60.7, 54.4, 50.5, 45.8, 41.5, 41.4, 39.5, 34.6, 34.4,32.0, 31.2, 29.8, 29.7, 29.4, 28.6, 26.1, 25.4, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C₉₀H₁₇₄N₃O₁₈P.H₂O: C, 66.10;H, 10.85; N, 2.57; P, 1.89.Found: C, 66.34;H, 10.69; N, 2.32; P, 1.99.

EXAMPLE 6 B5 PREPARATION OF3-HYDROXY-(S)-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]PROPYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₉H₁₉CO, X═Y═O, N=M=Q=0,R₄═R₅═R₇═R₉═H, R₆═OH, P=1 R₈═PO₃H₂).

(1) In the same manner as described in Example 4-(5), the compoundprepared in Example 4-(4) (2.0 g, 3.12 mmol) was acylated with(R)-3-decanoyloxytetradecanoic acid (1.36 g, 3.42 mmol) in the presenceof EDC·MeI (1.12 g, 3.74 mmol) and 4-pyrrolidinopyridine (100 mg) inCH₂Cl₂ (40 mL) to afford 2.49 g (79%) of3-benzyloxy-(S)-2-(allyloxycarbonylamino)propyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-decanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.7 Hz), 1.1-1.6(m, 34H), 1.36 (s, 3H), 1.46 (s, 3H), 2.27 (t, 2H, J=6.9 Hz), 248 (dd,1H, J=15.1, 6.0 Hz), 2.60 (dd, 1H, J=15.1, 6.7 Hz), 3.25 (m, 1H),3.35-4.0 (m, 9H), 4.23 (m, 1H), 4.42 (m, 1H), 4.52 (m, 4H), 4.95 (m,2H), 5.17 (m, 3H), 5.88 (m, 1H), 7.36 (m, 5H).

(2) In the same manner as described in Example 4-(6) the compoundprepared in (1) above (2.47 g, 2.42 mmol) was deprotected in TM (40 mL)in the presence of dimethyl malonate (2.0 mL, 1.75 mmol) andtetrakis(triphenylphosphine)palladium(0) (400 mg) and then acylated with(R)-3-decanoyloxytetradecanoic acid (1.06 g, 2.66 mmol) in the presenceof EEDQ (740 mg, 3 mmol) to afford 1.86 g (60%) of3-benzyloxy-(S)-2-[(R)-3-decanoyloxytetradecanoylamino]propyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-decanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.87 (t, 12H, J=6.7Hz), 1.1-1.7 (m, 68H), 1.37 (s, 3H), 1.46 (s, 3H), 2.32 (m, 4H), 2.50(dd, 1H, J=15.1, 6.0 Hz), 2.62 (dd, 1H, J=15.1, 6.8 Hz), 3.29 (m, 2H),3.44 (m, 1H), 3.55 (m, 1H), 3.74 (m, 3H), 3.93 (m, 1H), 4.18 (m, 1H),4.34 (m, 1H), 4.57 (d, 1H, J=11.8 Hz), 4.65 (m, 2H), 5.01 (m, 2H), 6.04(d, 1H, J=8.3 Hz), 7.36 (m, 5H).

(3) In the same manner as described in Example 4-(7) the compoundprepared in (2) above (900 mg, 0.72 mmol) was deprotected in 90% aqueousAcOH (40 mL), and then treated with pyridine (0.071 mL, 0.88 mmol) and2,2,2-trichloro-1,1-dimethylethyl chloroformnate (195 mg, 0.80 mmol) inCH₂Cl₂ followed by diphenyl chlorophosphate (0.23 mL, 1.10 mmol),triethylamine (0.20 mL, 1.46 mmol) and catalytic 4-pyrrolidinopyridine(50 mg) to afford 1.05 g (86%) of3-benzyloxy-(S)-2-[(R)-3-decanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.87 (t, 12H, J=6.3Hz), 1.1-1.6 (m, 68H), 1.78 (s, 3H), 1.86 (s, 3H), 2.01 (m, 1H), 2.18(m, 3H), 2.40 (m, 2H), 2.67 (m, 1H), 2.88 (d, 1H, J=6.5 Hz), 2.97 (d,1H, J=6.9 Hz), 3.41 (m, 2H), 3.72 (m, 1H), 3.82 (m, 1H), 4.24 (m, 1H),4.42 (d, 1H, J=11.8 Hz), 4.64 (m, 3H), 5.16 (m, 1H), 5.39 (m, 2H), 5.75(d, 1H, J=4.3 Hz), 6.05 (d, 1H, J=8.4 Hz), 7.22 (m, 15H).

(4) In the same manner as described in Example 2-(7), the compoundprepared in (3) above (1.0 g, 0.60 mmol) was deprotected with zinc (2.0g, 30 mmol) and acylated with (R)-3-decanoyloxytetradecanoic acid (285mg, 0.72 mmol) in the presence of EEDQ (210 mg, 0.86 mmol) to afford 332mg (34%) of3-benzyloxy-(S)-2-[(R)-3-decanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyltetradecanoyl]-β-D-glucopyranosideas a colorless amorphous solid.

(5) In the same manner as described in Example 2-(8), the compoundprepared in (4) above (332 mg, 0.20 mmol) was hydrogenated in thepresence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) andplatinum oxide (300 mg) in EtOH/AcOH (10:1) to afford 173 mg (55%) of3-hydroxy-(S)-2-[(R)-3-decanoyloxytetradecanoylamino]propyl2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as a white powder: mp 179-181° C.; IR (film) 3295,2956, 2923, 2853, 1732, 1650, 1555, 1467, 1377, 1320, 1169, 1134, 1104,1051, 979, 801, 721 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.9Hz), 1.1-1.7 (m, 111H), 2.2-2.7 (m, 12H), 3.07 (q, 6H, J=6.5 Hz),3.3-4.3 (m, 14H), 4.45 (d, 1 H, J=8.0 Hz), 5.0-5.3 (m, 4H), 7.39 (m,1H), 7.53 (d, 1H, J=9.1 Hz); ¹³C NMR (CDCl₃) δ 173.7, 173.4, 173.2,170.7, 170.5, 170.1, 101.0, 75.4, 73.1, 71.6, 71.1, 70.8, 70.5, 67.8,61.4, 60.8, 54.3, 50.4, 45.8, 41.3, 39.5, 34.5, 31.9, 29.8, 29.7, 29.4,25.4, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C₈₇H₁₆₈N₃O₁₈P.H₂O: C, 65.58;H, 10.75; N, 2.64; P, 1.94.Found: C, 65.49;H, 10.75; N, 2.64; P, 1.97.

EXAMPLE 7 B6 PREPARATION OF3-HYDROXY-(S)-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]PROPYL2-DEOXY-6-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-β-D-GlucopyranosideTriethylammonium Salt (Compound Of R₁═R₂═R₃═N—C₉H₁₉CO, X═Y═O, N=M=Q=0,R₄═R₅═R₇═R₈═H, R₆═OH, P=1, R₉═PO₃H₂).

(1) In the same manner as described in Example 4-(7) the compoundprepared in Example 6-(2) (900 mg, 0.72 mmol) was deprotected in 90%aqueous AcOH (20 mL). The residue was dissolved in CH₂Cl₂ (20 mL),cooled to 0° C., and treated with triethylamine (0.14 mL, 1.0 mmol) anddiphenyl chlorophosphate (0.17 mL, 0.8 mmol). The mixture was stirredfor an additional 6 h, and then quenched with 50 mL of 10% HCl. Theproduct was extracted with EtOAc (3×50 mL) and dried over Na₂SO₄.Chromatography on silica gel with 50% EtOAc/hexanes afforded 636 mg(63%) of 3-benzyloxy-(S)-2-[(R)-3-decanoyloxytetradecanoylamino]propyl2-deoxy-6-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.87 (t, 12H, J=6.0Hz), 1.1-1.6 (m, 68H), 1.79 (s, 3H), 1.86 (s, 3H), 2.01 (m, 1H), 2.18(m, 3H), 2.40 (m, 2H), 2.67 (m, 1H), 2.89 (d, 1H, J=6.5 Hz), 2.97 (d,1H, J=6.9 Hz), 3.41 (m, 2H), 3.75 (m, 1H), 3.82 (m, 1H), 4.24 (m, 1H),4.42 (d, 1H, J=11.8 Hz), 4.65 (m, 3H), 5.16 (m, 1H), 5.39 (m, 2H), 5.75(d, 1H, J=4.3 Hz), 6.05 (d, 1H, J=8.4 Hz), 7.22 (m, 15H).

(2) In the same manner as described in Example 2-(7), the compoundprepared in (1) above (620 g, 0.44 mmol) was deprotected with zinc (722mg, 11 mmol) and acylated with (R)-3-decanoyloxytetradecanoic acid (190mg, 0.48 mmol) in the presence of EEDQ (170 mg, 0.58 mmol) to afford 254mg (36%) of3-benzyloxy-(S)-2-[(R)-3-decanoyloxytetradecanoylamino]propyl2-deoxy-6-O-diphenylphosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyltetradecanoyl]-β-D-glucopyranosideas a colorless amorphous solid.

(3) In the same manner as described in Example 2-(8), the compoundprepared in (2) above (254 mg, 0.16 mmol) was hydrogenated in thepresence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) andplatinum oxide (300 mg) in EtOH/AcOH (10:1) to afford 34 mg (13%) of3-hydroxy-(S)-2-[(R)-3-decanoyloxytetradecanoylamino]propyl2-deoxy-6-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as a white powder: mp 169-171° C; IR (film) 3306,2922, 2853, 1732, 1644, 1548, 1467, 1377, 1316, 1165, 1106, 1053, 856,722 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.7 Hz), 1.1-1.7 (m,111H), 2.2-2.7 (m, 12H), 3.05 (m, 6H), 3.3-3.95 (m, 12H), 4.11 (m, 1H),4.34 (m, 1H), 4.89 (m, 1H), 5.0-5.3 (m, 4H). ¹³C NMR (CDCl₃) δ 173.8,173.4, 171.1, 170.5, 101.3, 75.3, 74.9, 71.2, 71.0, 70.6, 68.8, 67.3,65.1, 61.4, 53.4, 50.7, 45.9, 41.5, 41.3, 39.6, 34.6, 32.0, 29.8, 29.6,29.4, 25.3, 25.1, 22.7, 14.1, 8.7.

Anal. Calcd. for C₈₇H₁₆₈N₃O₁₈P.H₂O: C, 65.58;H, 10.75; N, 2.64; P, 1.94.Found: C, 65.60;H, 10.34; N, 2.36; P, 2.01.

EXAMPLE 8 B17 PREPARATION OF3-HYDROXY-(S)-2-[(R)-3-NONANOYLOXYTETRADECANOYLAMINO]PROPYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-NONANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-NONANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSIDETRIETYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₈H₁₇CO, X═Y═O, N═M═Q=O,R₄═R₅═R₇═R₉═H, R₆═OH, P=1, R₈═PO₃H₂).

(1) In the same manner as described in Example 4-(5), the compoundprepared in Example 4-(4) (1.0 g, 1.56 mmol) was acylated with(R)-3-nonanoyloxytetradecanoic acid (660 mg, 1.71 mmol) in the presenceof EDC·(560 mg, 1.87 mmol) and 4-pyrrolidinopyridine (50 mg) in CH₂Cl₂(20 mL) to afford 1.31 g (83%) of3-benzyloxy-(S)-2-(allyloxycarbonylamino)propyl2-deoxy-4,6-O-isopropylidene3-O-[(R)-3-nonanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.87 (t, 6H, J=6.8 Hz), 1.1-1.6(m, 32H), 1.37 (s, 3H), 1.46 (s, 3H), 2.27 (t, 2H, J=7.4 Hz), 2.50 (dd,1H, J=15.1, 6.0 Hz), 2.63 (dd, 1H, J=15.1, 6.8 Hz), 3.26 (m, 1H),3.35-4.0 (m, 9H), 4.32 (d, 1H, J=7.8 Hz), 4.41 (d, 1H, J=12.0 Hz), 4.51(m, 4H), 4.95 (m, 2H), 5.18 (m, 2H), 5.29 (d, 1H, J=17.2 Hz), 5.88 (m,1H), 7.36 (m, 5H).

(2) In the same manner as described in Example 4-(6) the compoundprepared in (1) above (1.29 g, 1.28 mmol) was deprotected in THF (20 mL)in the presence of dimethyl malonate (1.0 mL, 0.88 mmol) andtetrakis(triphenylphosphine)palladium(0) (200 mg) and then acylated with(R)-3-nonanoyloxytetradecanoic acid (540 mg, 1.41 mmol) in the presenceof EEDQ (370 mg, 1.5 mmol) to afford 1.02 g (65%) of3-benzyloxy-(S)-2-[(R)-3-nonanoyloxytetradecanoylamino]propyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-nonanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.87 (t, 12H, J=6.1Hz), 1.1-1.7 (m, 64H), 1.37 (s, 3H), 1.46 (s, 3H), 2.28 (m, 4H), 2.50(dd, 1H, J=15.5, 6.0 Hz), 2.62 (dd, 1H, J=14.8, 6.3 Hz), 3.27 (m, 2H),3.44 (m, 1H), 3.55 (m, 1H), 3.74 (m, 3H), 3.93 (m, 1H), 4.18 (m, 1H),4.34 (m, 2H), 4.57 (d, 1H, J=11.8 Hz), 4.65 (m, 2H), 4.97 (t, 1H, J=9.6Hz), 5.06 (d, 1H, J=8.6 Hz), 5.15 (m, 2H), 6.05 (d, 1H, J=8.2 Hz), 7.35(m, 5H).

(3) In the same manner as described in Example 4-(7) the compoundprepared in (2) above (1.0 g, 0.81 mmol) was deprotected in 90% aqueousAcOH (20 mL), treated with pyridine (0.080 mL, 0.98 mmol) and2,2,2-trichloro-1,1-dimethylethyl chloroformate (215 mg, 0.89 mmol) inCH₂Cl₂ followed by diphenyl chlorophosphate (0.25 mL, 1.22 mmol),triethylamine (0.21 mL, 1.52 mmol) and catalytic 4-pyrrolidinopyridine(50 mg) to afford 1.17 g (87%) of3-benzyloxy-(S)-2-[(R)-3-nonanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-nonanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.87 (t, 12H, J=6.1Hz), 1.1-1.6 (m, 64H), 1.78 (s, 3H), 1.86 (s, 3H), 2.01 (m, 1H), 2.18(m, 3H), 2.40 (m, 2H), 2.67 (m, 1H), 2.88 (d, 1H, J=6.5 Hz), 2.97 (d,1H, J=6.9 Hz), 3.41 (m, 2H), 3.72 (m, 1H), 3.82 (m, 1H), 4.24 (m, 1H),4.42 (d, 1H, J=11.8 Hz), 4.64 (m, 3H), 5.16 (m, 1H), 5.39 (m, 2H), 5.75(d, 1H, J=4.3 Hz), 6.05 (d, 1H, J=8.4 Hz), 7.22 (m, 15H).

(4) In the same manner as described in Example 2-(7), the compoundprepared in (3) above (1.1 g, 0.66 mmol) was deprotected with zinc (2.2g, 33 mmol) and acylated with (R)-3-nonanoyloxytetradecanoic acid (305mg, 0.79 mmol) in the presence of EEDQ (235 mg, 0.95 mmol) to afford 373mg (35%) of3-benzyloxy-(S)-2-[(R)-3-nonanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-2-[(R)-3-nonanoyloxytetradecanoylaminol-3-O-[(R)-3-nonanoyltetradecanoyl]-β-D-glucopyranosideas a colorless amorphous solid.

(5) In the same manner as described in Example 2-(8), the compoundprepared in (4) above (373 mg, 0.23 mmol) was hydrogenated in thepresence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) andplatinum oxide (300 mg) in EtOH/AcOH (10:1) to afford 43 mg (12%) of3-hydroxy-(S)-2-[(R)-3-nonanoyloxytetradecanoylamino]propyl2-deoxy-4-O-phosphono-2-[(R)-3-nonanoyloxytetradecanoylamino]-3-O-[(R)-3-nonanoyloxytetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as a white powder: mp 176-179° C.; IR (film) 3298,2956, 2923, 2853, 1733, 1646, 1551, 1467, 1337, 1316, 1254, 1166, 1106,1053, 722 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.87 (t, 18H, J=6.7 Hz), 1.1-1.7(m, 105H), 2.2-2.7 (m, 12H), 3.03 (q, 6H, J=7.0 Hz), 3.3-4.3 (m, 14H),4.43 (d, 1H, J=7.1 Hz), 5.0-5.3 (m, 4H), 7.12 (d, 1H, J=7.7 Hz), 7.17(d, 1H, J=8.2 Hz); ¹³C NMR (CDCl₃) δ 173.9, 173.5, 173.3, 170.8, 170.5,170.1, 100.9, 75.5, 73.1, 71.4, 71.1, 70.9, 70.6, 67.8, 61.6, 60.7,54.3, 50.5, 45.8, 41.6, 41.4, 39.5, 34.6, 34.4, 32.0, 31.9, 29.8, 29.4,29.3, 25.4, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C₈₈H₁₆₄N₃O₁₈P: C, 65.81;H, 10.65; N, 2.74; P, 2.02.Found: C, 66.14;H, 10.46; N, 2.58; P, 1.84.

EXAMPLE 9 B8 PREPARATION OF3-HYDROXY-(S)-2-[(R)-3-HEPTANOYLOXYTETRADECANOYLAMINO]PROPYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-HEPTANOYLOXYTETRADECAMINO]-3-O-[(R)-3-HEPTANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₆H₁₃CO, X═Y═O, N=M=Q=0,R₄═R₅═R₇═R₉═H, R₆═OH, P=1, R₈═PO₃H₂).

(1) In the same manner as described in Example 4-(5), the compoundprepared in Example 4-(4) (1.0 g, 1.56 mmol) was acylated with(R)-3-heptanoyloxytetradecanoic acid (610 mg, 1.71 mmol) in the presenceof EDC·MeI (560 mg, 1.87 mmol) and 4-pyrrolidinopyridine (50 mg) inCH₂Cl₂ (20 mL) to afford 1.24 g (82%) of3-benzyloxy-(S)-2-(allyloxycarbonylamino)propyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-heptanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.0 Hz), 1.1-1.6(m, 28H), 1.38 (s, 3H), 1.47 (s, 3H), 2.29 (t, 2H, J=7.4 Hz), 2.51 (dd,1H, J=15.1, 6.0 Hz), 2.63 (dd, 1H, J=15.1, 6.8 Hz), 3.26 (m, 1H),3.35-4.0 (m, 9H), 4.32 (d, 1H, J=7.3 Hz), 4.41 (d, 1H, J=12.0 Hz), 4.51(m, 4H), 4.95 (m, 2H), 5.18 (m, 2H), 5.29 (d, 1H, J=17.3 Hz), 5.88 (m,1H), 7.36 (m, 5H).

(2) In the same manner as described in Example 4-(6) the compoundprepared in (1) above (1.22 g, 1.25 mmol) was deprotected in TBF (20 mL)in the presence of dimethyl malonate (1.0 mL, 0.88 mmol) andtetrakis(triphenylphosphine)palladium(0) (200 mg) and then acylated with(R)-3-heptanoyloxytetradecanoic acid (490 mg, 1.38 mmol) in the presenceof EEDQ (370 mg, 1.5 mmol) to afford 925 mg (62%) of3-benzyloxy-(S)-2-[(R)-3-heptanoyloxytetradecanoylamino]propyl2-deoxy4,6-O-isopropylidene-3-O-[(R)-3-heptanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.87 (t, 12H, J=6.7Hz), 1.1-1.7 (m, 56H), 1.37 (s, 3H), 1.46 (s, 3H), 2.32 (m, 4H), 2.50(dd, 1H, J=15.1, 6.0 Hz), 2.62 (dd, 1H, J=15.1, 6.8 Hz), 3.29 (m, 2H),3.44 (m, 1H), 3.55 (m, 1H), 3.74 (m, 3H), 3.93 (m, 1H), 4.18 (m, 1H),4.34 (m, 1H), 4.57 (d, 1H, J=11.8 Hz), 4.65 (m, 2H), 5.01 (m, 2H), 6.04(d, 1H, J=8.3 Hz), 7.36 (m, 5H).

(3) In the same manner as described in Example 4-(7) the compoundprepared in (2) above (920 mg, 0.76 mmol) was deprotected in 90% aqueousAcOH (20 mL), and then treated with pyridine (0.075 mL, 0.92 mmol) and2,2,2-trichloro-1,1-dimethylethyl chloroformate (200 mg, 0.84 mmol) inCH₂Cl₂ followed by diphenyl chlorophosphate(0.24 mL, 1.14 mmol),triethylamine (0.21 mL, 1.52 mmol) and catalytic 4-pyrrolidinopyridine(50 mg) to afford 1.03 g (83%) of3-benzyloxy-(S)-2-[(R)-3-heptanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-heptanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.87 (t, 12H, J=6.3Hz), 1.1-1.6 (m, 56H), 1.78 (s, 3H), 1.86 (s, 3H), 2.01 (m, 1H), 2.18(m, 3H), 2.40 (m, 2H), 2.67 (m, 1H), 2.88 (d, 1H, J=6.5 Hz), 2.97 (d,1H, J=6.9 Hz), 3.41 (m, 2H), 3.72 (m, 1H), 3.82 (m, 1H), 4.24 (m, 1H),4.42 (d, 1H, J=11.8 Hz), 4.64 (m, 3H), 5.16 (m, 1H), 5.39 (m, 2H), 5.75(d, 1H, J=4.3 Hz), 6.05 (d, 1H, J=8.4 Hz), 7.22 (m, 15H).

(4) In the same manner as described in Example 2-(7), the compoundprepared in (3) above (1.0 g, 0.61 mmol) was deprotected with zinc (2.0g, 31 mmol) and acylated with (R)-3-heptanoyloxytetradecanoic acid (260mg, 0.73 mmol) in the presence of EEDQ (220 mg, 0.88 mmol) to afford 203mg (21%) of3-benzyloxy-(S)-2-[(R)-3-heptanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-2-[(R)-3-heptanoyloxytetradecanoylamino]-3-O-[(R)-3-heptanoyloxytetradecanoyl]-β-D-glucopyranosideas a colorless amorphous solid.

(5) In the same manner as described in Example 2-(8), the compoundprepared in (4) above (203 mg, 0.13 mmol) was hydrogenated in thepresence of palladium hydroxide (100 mg) on carbon in EtOH (10 mL) andplatinum oxide (200 mg) in EtOH/AcOH (10:1) to afford 39 mg (21%) of3-hydroxy-(S)-2-[(R)-3-heptanoyloxytetradecanoylamino]propyl2-deoxy-4-O-phosphono-2-[(R)-3-heptanoyloxytetradecanoylaminol-3-O-[(R)-3-heptanoyloxytetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as a white powder: mp 171-172° C.; IR (film) 3305,2955, 2924, 2853, 1734, 1644, 1553, 1466, 1377, 1170, 1102, 1052, 722cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (m, 18H), 1.1-1.7 (m, 93H), 2.2-2.7(m, 12H), 3.06 (q, 6H, J=7.1 Hz), 3.3-4.0 (m, 13H), 4.23 (q, 1H, J=9.3Hz), 4.43 (d, 1H, J=8.2 Hz), 5.0-5.3 (m, 4H), 7.30 (d, 1H, J=8.5 Hz),7.43 (d, 1H, J=8.5 Hz); ¹³C NMR (CDCl₃) δ 173.8, 173.5, 173.2, 170.8,170.5, 170.2, 101.0, 77.2, 75.5, 73.1, 71.6, 71.1, 70.9, 70.6, 67.8,61.6, 60.8, 54.4, 50.5, 45.8, 41.6, 41.4, 39.5, 34.6, 34.4, 32.0, 31.6,29.8, 29.6, 29.4, 28.9, 25.4, 25.1, 22.7, 22.6, 14.1, 8.6.

Anal. Calcd. for C₇₈H₁₅₀N₃O₁₈P.H₂O: C, 63.86;H, 10.44; N, 2.86; P, 2.11.Found: C, 63.47;H, 10.20; N, 2.59; P, 2.02.

EXAMPLE 10 B9 PREPARATION OF4-HYDROXY-(S)-3-[(R)-3-DECANOYLOXYTETRADECANOYL]BUTYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLTETRADECANOYL]-62-D-GLUCOPYRANOSIDE TRIETHYLAMMONIUM SALT (COMPOUND (I),R₁═R₂═R₃═N—C₉H₁₉CO, X═Y═O, N=P=1, M=Q=0, R₄═R₅═R₇═R₉═H, R₆═OH,R₈═PO₃H₂).

(1) In the same manner as described in Example 4-(3) the compoundprepared in Example 4-(1) (3.1 g, 5.9 mmol) and(R)-3-(allyloxycarbonylamino)-4-benzyloxy-1-butanol (1.1 g, 3.94 mmol)were coupled in the presence of boron trifluoride etherate (3.0 mL, 23.6mmol) to afford 1.96 g (67%) of4-benzyloxy-(S)-3-(allyloxycarbonylamino)butyl2-deoxy-3,4,6-tri-O-acetyl-2-(2,2,2-trichloroethoxycarbonylamnino)-β-D-glucopyranosideas an amorphous solid. In the same manner as described in Example 4-(4)the compound prepared above (1.8 g, 2.43 mmol) was deacylated inmethanol (25 mL) with ammonium hydroxide (5 mL) and then treated with2,2-dimethoxypropane (25 mL) and camphorsulfonic acid (100 mg) to afford1.34 g (84%) of 4-benzyloxy-(S)-3-(allyloxycarbonylamino)butyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranoside.

(2) In the same manner as described in Example 4-(5), the compoundprepared in (1) above (1.0 g, 1.53 mmol) was acylated with(R)-3-decanoyloxytetradecanoic acid (670 mg, 1.68 mmol) in the presenceof EDC·MeI (550 mg, 1.85 mmol) and 4-pyrrolidinopyridine (50 mg) inCH₂Cl₂ (15 mL) to afford 1.03 g (65%) of4-benzyloxy-(S)-3-(allyloxycarbonylamino)butyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-decanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.9 Hz), 1.1-1.6(m, 34H), 1.37 (s, 3H), 1.47 (s, 3H), 1.85 (m, 2H), 2.28 (t, 2H, J=7.6Hz), 2.50 (dd, 1H, J=15.1, 6.0 Hz), 2.63 (dd, 1H, J=15.1, 6.7 Hz), 3.30(m, 1H), 3.49 (m, 4H), 3.68 (t, 1H, J=9.4 Hz), 3.77 (t, 1H, J=10.4 Hz),3.92 (m, 3H), 4.54 (m, 5H), 4.69 (m, 2H), 5.1-5.4 (m, 4H), 5.91 (m, 1H),7.33 (m, 5H).

(3) In the same manner as described in Example 4-(6) the compoundprepared in (2) above (1.0 g, 0.97 mmol) was deprotected in THF (20 mL)in the presence of dimethyl malonate (1.0 mL, 0.88 mmol) andtetrakis(triphenylphosphine)palladium(0) (200 mg) and then acylated with(R)-3-decanoyloxytetradecanoic acid (425 mg, 1.07 mmol) in the presenceof EEDQ (317 mg, 1.28 mmol) to afford 660 mg (51%) of4-benzyloxy-(S)-3-[(R)-3-decanoyloxytetradecanoylamino]propyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-decanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.6Hz), 1.1-1.7 (m, 68H), 1.37 (s, 3H), 1.47 (s, 3H), 2.26 (q, 2H, J=7.1Hz), 2.41 (m, 2H), 2.62 (dd, 1H, J=14.9, 6.4 Hz), 3.29 (m, 1H), 3.48 (m,3H), 3.71 (m, 2H), 3.92 (m, 2H), 4.18 (m, 1H), 4.49 (m, 2H), 4.68 (q,2H, J=11.5 Hz), 5.15 (m, 2H), 5.55 (d, 1H, J=8.8 Hz), 6.17 (d, 1H, J=7.2Hz), 7.32 (m, 5H).

(4) In the same manner as described in Example 4-(7) the compoundprepared in (3) above (640 mg, 0.48 mmol) was deprotected in 90% aqueousAcOH (20 mL), and then treated with pyridine (0.047 mL, 0.58 mmol) and2,2,2-trichloro-1,1-dimethylethyl chloroformate (127 mg, 0.53 mmol) inCH₂Cl₂ followed by diphenyl chlorophosphate (0.15 mL, 0.72 mmol),triethylamine (0.13 mL, 0.96 mmol) and catalytic 4-pyrrolidinopyridine(50 mg) to afford 389 mg (47%) of4-benzyloxy-(S)-3-[(R)-3-decanoyloxytetradecanoyl]butyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.6Hz), 1.1-1.6 (m, 68H), 1.79 (s, 3H), 1.86 (s, 3H), 2.22 (m, 4H), 2.40(m, 4H), 3.49 (m, 4H), 3.78 (m, 1H), 3.93 (m, 1H), 4.1-4.5 (m, 5H),4.9-4.6 (m, 4H), 5.13 (m, 2H), 5.51 (t, 1H, J=8.9 Hz), 5.84 (d, 1H,J=6.9 Hz), 6.09 (d, 1H, J=8.0 Hz), 7.26 (m, 15H).

(5) In the same manner as described in Example 2-(7), the compoundprepared in (4) above (375 g, 0.23 mmol) was deprotected with zinc (752mg, 11.5 mmol) and acylated with (R)-3-decanoyloxytetradecanoic acid(101 mg, 0.25 mmol) in the presence of EEDQ (70 mg, 0.28 mmol) to afford270 mg (67%) of 4-benzyloxy-(S)-3-[(R)-3-decanoyloxytetradecanoyl]butyl2-deoxy-4-O-diphenylphosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyltetradecanoyl]-β-D-glucopyranosideas a colorless amorphous solid.

(6) In the same manner as described in Example 2-(8), the compoundprepared in (5) above (270 mg, 0.15 mmol) was hydrogenated in thepresence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) andplatinum oxide (300 mg) in EtOH/AcOH (10:1) to afford 93 mg (39%) of4-hydroxy-(S)-3-[(R)-3-decanoyloxytetradecanoyl]butyl2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyltetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as a white powder: mp 179-181° C. (dec): IR (film)3287, 2956, 2923, 2853, 1734, 1654, 1552, 1466, 1378, 1246, 1164, 1106,1085, 1052, 721 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.9 Hz),1.1-1.7 (m, 111H), 2.2-2.7 (m, 14H), 3.06 (q, 6H, J=6.9 Hz), 3.2-4.0 (m,13H), 4.21 (m, 1H), 4.50 (d, 1H, J=7.7 Hz), 5.0-5.3 (m, 4H), 7.11 (m,2H); 1³C NMR (CDCl₃) δ 173.8, 173.5, 173.3, 170.9, 170.5, 170.1, 101.1,77.2, 75.5, 72.8, 71.3, 71.0, 70.6, 66.4, 64.0, 60.7, 54.8, 50.2, 45.8,41.6, 39.5, 34.6, 34.5, 34.4, 32.0, 30.6, 29.8, 29.7, 29.6, 29.5, 29.4,25.4, 25.1, 22.7, 14.2, 8.6.

Anal. Calcd. for C₁₈H₁₇₀N₃O₁₈P: C, 66.65;H, 10.78; N, 2.64; P, 1.95.Found: C, 66.65 H, 10.68; N, 2.50; P, 1.94.

EXAMPLE 11 B10 PREPARATION OF4-HYDROXY-(S)-2-[(R)-3-DECANOYLOXYTETRADECANOYL]BUTYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLTETRADECANOYL]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₉H₁₉CO, X═Y═O, N=M=Q=0,R₄═R₅═R₇═R₉═H, R₆═OH, P=2, R₈═PO₃H₂).

(1) In the same manner as described in Example 4-(3) the compoundprepared in Example 4-(1) (5.1 g, 9.7 mmol) and(R)-2-(allyloxycarbonylamino)-4-benzyloxy-1-butanol (1.8 g, 6.45 mmol)were coupled in the presence of boron trifluoride etherate (4.9 mL, 38.0mmol) to afford 2.92 g (61%) of4-benzyloxy-(S)-2-(allyloxycarbonylamino)propyl2-deoxy-3,4,6-tri-O-acetyl-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid. In the same manner as described in Example 4-(4)the compound prepared above (2.6 g, 3.51 mmol) was deacylated inmethanol (35 mL) with ammonium hydroxide (7 mL) and then treated with2,2-dimethoxypropane (35 mL) and camphorsulfonic acid (100 mg) to afford1.9 g (72%) of 4-benzyloxy-(S)-2-(allyloxycarbonylamino)butyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranoside.

(2) In the same manner as described in Example 4-(5), the compoundprepared in (1) above (1.0 g, 1.53 mmol) was acylated with(R)-3-decanoyloxytetradecanoic acid (670 mg, 1.68 mmol) in the presenceof EDC·MeI (550 mg, 1.85 mmol) and 4-pyrrolidinopyridine (50 mg) inCH₂Cl₂ (15 mL) to afford 1.28 g (81%) of4-benzyloxy-(S)-2-(allyloxycarbonylamino)butyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-decanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.9 Hz), 1.1-1.7(m, 34H), 1.37 (s, 3H), 1.47 (s, 3H), 1.82 (m, 2H), 2.28 (t, 2H, J=7.7Hz), 2.50 (dd, 1H, J=15.3, 6.0 Hz), 2.63 (dd, 1H, J=15.2, 6.7 Hz), 3.16(m, 1H), 3.56 (m, 3H), 3.65 (t, 1H, J=9.6 Hz), 3.75 (t, 1H, J=10.4 Hz),3.88 (m, 4H), 4.32 (d, 1H, J=8.5 Hz), 4.46 (s, 2H), 4.54 (m, 2H), 4.67(m, 2H), 4.90 (m, 11H), 5.26 (m, 3H), 5.89 (m, 1H), 7.33 (m, 5H).

(3) In the same manner as described in Example 4-(6) the compoundprepared in (2) above (1.25 g, 1.21 mmol) was deprotected in THF (20 mL)in the presence of dimethyl malonate (1.0 mL, 0.88 mmol) andtetrakis(triphenylphosphine)palladium(0) (200 mg) and then acylated with(R)-3-decanoyloxytetradecanoic acid (530 mg, 1.33 mmol) in the presenceof EEDQ (362 mg, 1.46 mmol) to afford 1.16 g (72%) of4-benzyloxy-(S)-3-[(R)-3-decanoyloxytetradecanoylamino]propyl2-deoxy-4,6-O-isopropylidene-3-O-[(R)-3-decanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-13-D-glucopyranosideas a colorless amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.4Hz), 1.1-1.7 (m, 68H), 1.37 (s, 3H), 1.45 (s, 3H), 2.26 (q, 2H, J=7.4Hz), 2.34 (m, 1H), 2.50 (dd, 1H, J=15.1, 6.0 Hz), 2.62 (dd, 1H, J=15.4,6.3 Hz), 3.12 (m, 1H), 3.5-3.95 (m, 7H), 4.14 (m, 1H), 4.29 (d, 1H,J=8.0 Hz), 4.67 (m, 2H), 4.86 (t, 1H, J=9.6 Hz),5.15 (m, 2H), 6.16 (d,1H, J=8.3 Hz), 7.35 (m, 5H).

(4) In the same manner as described in Example 4-(7) the compoundprepared in (3) above (1.1 g, 0.83 mmol) was deprotected in 90% aqueousAcOH (20 mL), and then treated with pyridine (0.080 mL, 1.0 mmol) and2,2,2-trichloro-1,1-dimethylethyl chloroformate (220 mg, 0.91 mmol) inCH₂Cl₂ followed by diphenyl chlorophosphate (0.26 mL, 1.25 mmol),triethylamine (0.23 mL, 1.66 mmol) and catalytic 4-pyrrolidinopyridine(50 mg) to afford 802 mg (56%) of4-benzyloxy-(S)-2-[(R)-3-decanoyloxytetradecanoyl]butyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas a colorless amorphous solid: ¹HNMR (CDCl₃) δ 0.87 (t, 12H, J=6.8 Hz),1.1-1.6 (m, 68H), 1.79 (s, 3H), 1.88 (s, 3H), 2.23 (m, 4H), 2.37 (m,4H), 3.57 (m, 4H), 3.83 (m, 1H), 4.29 (m, 3H), 4.44 (m, 2H), 4.69 (m,4H), 5.14 (m, 4H), 5.62 (d, 1H, J=7.6 Hz), 6.15 (d, 1H, J=8.3 Hz), 7.25(m, 15H).

(5) In the same manner as described in Example 2-(7), the compoundprepared in (4) above (750 mg, 0.43 mmol) was deprotected with zinc(1.42 g, 21.7 mmol) and acylated with (R)-3-decanoyloxytetradecanoicacid (190 mg, 0.48 mmol) in the presence of EEDQ (130 mg, 0.53 mmol) toafford 483 mg (64%) of4-benzyloxy-(S)-2-[(R)-3-decanoyloxytetradecanoyl]butyl2-deoxy-4-O-diphenylphosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyltetradecanoyl]-β-D-glucopyranosideas a colorless amorphous solid.

(6) In the same manner as described in Example 2-(8), the compoundprepared in (5) above (483 mg, 0.27 mmol) was hydrogenated in thepresence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) andplatinum oxide (300 mg) in EtOH/AcOH (10:1) to afford 238 mg (55%) of4-hydroxy-(S)-2-[(R)-3-decanoyloxytetradecanoyl]butyl2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyltetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as a white powder: mp 181-183° C. (dec): IR (film)3294, 2956, 2923, 2853, 1732, 1650, 1556, 1466, 1377, 1320, 1246, 1172,1108, 1082, 1058, 859, 721 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H,J=6.9 Hz), 1.1-1.7 (m, 111H), 2.2-2.7 (m, 14H), 3.06 (q, 6H, J=7.1 Hz),3.2-4.0 (m, 13H), 4.21 (m, 1H), 4.46 (d, 1H, J=8.3 Hz), 5.0-5.3 (m, 4H);¹³C NMR (CDCl₃) δ 173.9, 173.4, 173.2, 171.2, 170.7, 101.0, 77.2, 75.4,73.1, 71.4, 71.3, 71.1, 70.9, 70.6, 60.7, 58.4, 54.7, 46.3, 45.9, 41.6,41.1, 39.7, 34.8, 34.6, 34.4, 31.9, 29.8, 29.6, 29.5, 29.3, 25.4, 25.3,25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C₈₈H₁₇₀N₃O₁₈P: C, 66.51;H, 10.78; N, 2.64; P, 1.95.Found: C, 66.81;H, 10.68; N, 2.53; P, 1.79.

EXAMPLE 12 B11 PREPARATION OFN-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-TETRADECANOYLOXYNETRADECANOYLAMINO]-3-O-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₃H₂₇CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

(1) In the same manner as described in Example 2-(5), L-serine benzylester (0.212 g, 1.08 mmol) was acylated with(R)-3-tetradecanoyloxytetradecanoic acid (0.541 g, 1.19 mmol) in thepresence of EDC·MeI (0.353 g, 1.19 mmol) to give 0.642 g (94%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-L-serine benzyl ester as a waxysolid: mp 56-61° C.; 1H NMR (CDCl₃) δ 0.88 (t, 6H, J=7 Hz), 1.1-1.7 (m,42H), 2.29 (t, 2H, J=7.5 Hz), 2.50 (m, 2H), 3.87 (br t, 1H), 3.95 (m,2H), 4.65 (m, 1H), 5.1-5.25 (m, 3H), 6.69 (d, 1H, J=7 Hz), 7.34 (br s,5H).

(2) In the same manner as described in Example 2-(6), the compoundprepared in (1) above (0.19 g, 0.30 mmol) and the compound prepared inExample 2-(4) (0.635 g, 0.478 mmol) were coupled in the presence ofmercury cyanide (0.3 g , 1.2 mmol) to give 0.425 g (77%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosyl]-L-serinebenzyl ester as an amorphous solid.

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (0.405 g, 0.22 mmol) was deprotected with zinc(0.72 g, 11 mmol) and acylated with (R)-3-tetradecanoyloxytetradecanoicacid (0.12 g, 0.26 mmol) in the presence of EEDQ (0.082 g, 0.33 mmol) togive 0.277 g (66%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-2-[(R)-3-tetradecanoyloxytetradecanoylamino]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinebenzyl ester as an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 18H, J=6.5Hz) 1.0-1.75 (m, 126H), 2.15-2.45 (m, 10H), 2.53 (dd, 1H, J=14.7, 6.0Hz), 2.67 (dd, 1H, J=14, 6.0 Hz), 3.25 (br t, 1H, J=7 Hz), 3.35-3.75 (m,4H), 3.88 dd, 1H, J=11.1 Hz), 4.23 dd, 1H, J=11.1, 3 Hz), 4.6-4.75 (m,2H), 5.03 (d, 1H, J=8.1 Hz), 5.05-5.25 (m, 4H), 5.48 (t, 1H, J=˜10 Hz),6.40 (d, 1H, J=7.5 Hz), 7.01 d, 1H, J=8.1 Hz), 7.1-7.4 (m, 15H).

(4) In the same manner as described in Example 2-(8), the compoundrepared in (3) above (0.253 g, 0.133 mmol) was hydrogenated in thepresence of 5% alladium on carbon (50 mg) and platinum oxide (120 mg) togive 0.155 g (62%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-tetradecanoyloxytetradecanoylamnino]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinetriethylammonium salt as a colorless solid: mp 180° C. (dec); IR (film)3322, 2956, 2924, 2852, 1736, 1732, 1681, 1673, 1667, 1660, 1651, 1467,1456, 1247, 1174, 1110, 1081 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H,J=˜7 Hz), 1.0-1.7 (m, 135H), 2.2-2.75 (m, 12H), 3.05 (q, 6H, J=7 Hz),3.30 (br s, 13H), 3.7-3.9 (m, 3H), 3.96 (d, 1H, J=12 Hz), 4.05-4.3 (m,2H), 4.34 (m, 1H), 4.53 (d, 1H, J=7.8 Hz), 5.05-5.3 (m, 4H), 7.25-7.35(m, 2H); ¹³C NMR (CDCl₃) δ 173.4, 173.2, 171.0, 170.3, 170.2, 169.9,169.8, 100.8, 75.1, 73.4, 71.1, 70.7, 70.4, 70.3, 60.2, 54.3, 45.6,41.2, 41.1, 39.2, 34.6, 34.4, 34.2, 32.0, 29.8, 29.5, 25.4, 25.2, 22.7,14.2, 8.6.

Anal. Calcd for C₉₉H₁₉₀N₃O₁₉P.5H₂O: C, 64.35;H, 10.91; N, 2.27; P, 1.68.Found: C, 64.16;H, 10.92; N, 2.37; P, 1.91.

EXAMPLE 13 B12 PREPARATION OFN-[(R)-3-DODECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DODECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DODECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₁H₂₃CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

(1) In the same manner as described in Example 2-(5), L-serine benzylester (390 mg, 2.0 mmol) was acylated with(R)-3-dodecanoyloxytetradecanoic acid (935 mg, 2.2 mmol) in the presenceof EDC·MeI (745 mg, 2.5 mmol) in CH₂C₂ to afford 1.08 g (90%) ofN-[(R)-3-dodecanoyloxytetradecanoyl]-L-serine benzyl ester: mp 53-54° C.¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.5 Hz), 1.1-1.6 (m, 46H), 2.30 (t, 2H,J=7.7 Hz), 2.50 (d, 2H, 5.6 Hz), 2.62 (t, 1H, J=6.2 Hz), 3.97 (m, 2H),4.65 (m, 1H), 5.19 (m, 3H), 6.63 (d, 1H, J=6.8 Hz), 7.35 (br s, 5H).

(2) In the same manner as described in Example 2-(2), the compoundprepared in Example 2-(1) (1.0 g, 2.02 mmol) was acylated with(R)-3-dodecanoyloxytetradecanoic acid (946 mg, 2.22 mmol) in thepresence of EDC·MeI (720 mg, 2.4 mmol) and 4-pyrrolidinopyridine (100mg) in CH₂Cl₂, and then deprotected in aqueous AcOH (25 mL) to afford1.30 g (81%) of 2-(trimethylsilyl)ethyl2-deoxy-3-O-[(R)-3-dodecanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.00 (s, 9H), 0.88 (m, 8H), 1.25(m, 28H), 1.59 (m, 4H), 2.30 (t, 2H, J=7.5 Hz), 2.52 (m, 2H), 3.42 (m,1H), 3.55 (m, 1H), 3.66 (m, 1H), 3.83 (dd, 1H, J=11.8, 4.6 Hz), 3.94 (m,2H), 4.57 (d, 1H, J=8.2 Hz), 4.71 (m, 2H), 5.07 (m, 2H), 5.27 (d, 1H,J=8.8 Hz).

(3) In the same manner as described in Example 2-(3), the compoundprepared in (2) above (1.30 g, 1.51 mmol) was treated with2,2,2-trichloro-1,1-dimethylethyl chloroformate (398 mg, 1.66 mmol) andpyridine (0.15 mL, 1.83 mmol) in CH₂Cl₂ (25 mL) followed bytriethylamine (0.42 mL, 3.02 mmol), diphenyl chlorophosphate (0.47 mL,2.27 mmol) and 4-pyrrolidinopyridine (100 mg) to afford 1.39 g (71%) of2-(trimethylsilyl)ethyl2-deoxy-4O-diphenylphosphono-3-O-[(R)-3-dodecanoyloxytetradecanoyl]-6O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.0 (s, 9H), 0.88 (m, 8H),1.1-1.7 (m, 46H), 1.77 (s, 3H), 1.85 (s, 3H), 2.23 (m, 6H), 3.34 (m,1H), 3.59 (m, 1H), 3.80 (m, 1H), 3.96 (m, 1H), 4.32 (m, 2 H), 4.63 (m,2H), 4.83 (d, 1H, J=11.9 Hz), 5.02 (d, 1H, J=8.2 Hz), 5.20 (m, 1H), 5.65(m, 2H), 7.29 (m, 10H).

(4) The compound prepared in (3) above (1.30 g, 1.0 mmol) in CH₂Cl₂ (15mL) was treated at 0° C. with TFA (5 mL) and then allowed to warm toroom temperature for 18 h. The solvent was removed in vacuo and theremaining TFA was removed by azeotroping with toluene. The lactol wastreated with the Vilsmeier reagent prepared from DMF (0.39 mL, 5.0 mmol)and oxalyl chloride (0.22 mL, 2.5 mmol) in CH₂Cl₂ (20 mL) at 0° C. Thereaction was allowed to warm slowly to room temperature overnight andwas partitioned between 50 mL of saturated aqueous NaHCO₃ and ether (50mL). The layers were separated and the organic phase was dried overNa₂SO₄ and concentrated in vacuo. Purification by flash chromatographyon silica gel with 10% EtOAc/hexanes afforded 1.09 g (90%) of2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-dodecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosylchloride as a white foam: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.8 Hz),1.2-1.70 (m, 46H), 1.78 (s, 3H), 1.88 (s, 3H), 2.18 (t, 2H, J=7.7 Hz),2.43 (m, 2H), 4.30 (m, 4 H), 4.72 (m, 3H), 5.09 (m, H), 5.50 (t, 1H,J=9.5 Hz), 5.79 (d, 1H, J=8.0 Hz), 6.27 (d, 1H, J=3.6 Hz), 7.19 (m,10H).

(5) To a solution of compounds prepared in (1) and (4) (540 mg, 0.90mmol, and 1.0 g, 0.82 mmol, respectively) in 1,2-dichloroethane (20 mL),powdered 4A molecular sieves (300 mg) were added and the suspension wasstirred for 30 min. AgOTf (1.16 g, 4.5 mmol) was added in one portion,after 30 min the slurry was filtered through silica gel and eluted with30% EtOAc/hexanes to afford 1.10 g (75%) ofN-[(R)-3-dodecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-dodecanoyloxytetradecanoyl]-6O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosyl]-L-serinebenzyl ester: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.5 Hz), 1.1-1.65 (m,92H), 1.77 (s, 3H), 1.85 (s, 3H), 2.1-2.5 (mn, 8H), 3.67 (m, 2H), 4.30(m, 3H), 4.72 (m, 5H), 5.18 (m, 4H), 5.46 (m, 1H), 6.07 (m, 1H), 6.62(d, 1H, J=7.9 Hz), 7.05-7.45 (m, 15H).

(6) In the same manner as described in Example 2-(7), the compoundprepared in (5) above (1.0 g, 0.56 mmol) was deprotected with zinc (1.83g, 28 mmol) and acylated with (R)-3-dodecanoyloxytetradecanoic acid (285mg, 0.67 mmol) in the presence of EEDQ (185 mg, 0.74 mmol) to afford 420mg (44%) ofN-[(R)-3-dodecanoyloxytetradecanoyl)-O-[2-deoxy-4-O-diphenylphosphono-2-[(R)-3-dodecanoyloxytetradecanoylamino]-3-O-[(R)-3-dodecanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinebenzyl ester as an amorphous solid.

(7) In the same manner as described in Example 2-(8), the compoundprepared in (6) above (420 mg, 0.24 mmol) was hydrogenated in thepresence of palladium hydroxide on carbon in EtOH (10 mL) and platinumoxide (400 mg) in EtOH/AcOH (10:1) to afford 240 mg (60%) ofN-[(R)-3-dodecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-dodecanoyloxytetradecanoylamino]-3-O-[(R)-3-dodecanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinetriethylammonium salt as a white powder: mp 181-182° C.; IR (film) 3289,2956, 2920, 2851, 1731, 1656, 1557, 1467, 1378, 1182, 1108, 1080, 1052,852, 721 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.7 Hz), 1.1-1.7(m, 123H), 2.2-2.7 (m, 12H), 3.06 (q, 6H, J=7.2 Hz), 3.35 (m, 1H), 3.70(m, 6H), 3.88 (m, 2H), 4.20 (m, 1H), 4.56 (d, 1H, J=8.1 Hz), 4.59 (br s,1H), 5.16 (m, 4H); ¹³C NMR (CDCl₃) δ 176.9, 173.3, 173.2, 172.7, 169.6,169.1, 101.5, 74.8, 71.2, 70.9, 69.2, 60.5, 53.1, 51.4, 46.1, 41.5,41.0, 39.2, 34.3, 34.2, 34.0, 32.0, 29.8, 29.7, 29.4, 29.2, 25.6, 25.3,25.2, 25.1, 22.7, 14.1, 8.7.

Anal. Calcd. for C₉₃H₁₇₈N₃O₁₉P.H₂O: C, 66.04;H, 10.73; N, 2.48; P, 1.83.Found: C, 66.04;H, 10.73; N, 2.48; P, 1.86.

EXAMPLE 14 B13 PREPARATION OFN-[(R)-3-UNDECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-UNDECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-UNDECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N-C₁₀H₂₁CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

(1) In the same manner as described in Example 2-(5), L-serine benzylester (390 mg, 2.0 mmol) was acylated with(R)-3-undecanoyloxytetradecanoic acid (905 mg, 2.2 mmol) in the presenceof EDC·MeI (745 mg, 2.5 mmol) in CH₂Cl₂ to afford 1.08 g (92%) ofN-[(R)-3-undecanoyloxytetradecanoyl]-L-serine benzyl ester: mp 53-54°C.; ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.9 Hz), 1.1-1.7 (m, 44H), 2.30 (t,2H, J=7.7 Hz), 2.49 (d, 2H, J=5.8 Hz), 3.99 (m, 2H), 4.65 (m, 1H), 5.19(m, 3H), 6.58 (d, 1H, J=6.9 Hz), 7.35 (br s, 5H).

(2) In the same manner as described in Example 2-(2), the compoundprepared in Example 2-(1) (1.0 g, 2.02 mmol) was acylated with(R)-3-undecanoyloxytetradecanoic acid (915 mg, 2.22 mmnol) in thepresence of EDC·MeI (720 mg, 2.4 mmol) and 4-pyrrolidinopyridine (100mg) in CH₂Cl₂, and then deprotected in aqueous AcOH (25 mL) to afford1.41 g (82%) of 2-(trimethylsilyl)ethyl2-deoxy-3-O-[(R)-3-undecanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.00 (s, 9H), 0.88 (m, 8H), 1.25(m, 32H), 1.60 (m, 4H), 2.31 (t, 2H, J=7.5 Hz), 2.52 (m, 2H), 3.42 (m,1H), 3.55 (m, 1H), 3.66 (m, 1H), 3.83 (dd, 1H, J=11.8, 4.6 Hz), 3.94 (m,2H), 4.57 (d, 1H, J=8.2 Hz), 4.71 (m, 2H), 5.07 (m, 2H), 5.27 (d, 1H,J=8.7 Hz).

(3) In the same manner as described in Example 2-(3), the compoundprepared in (2) above (1.30, 1.53 mmol) was treated with2,2,2-trichloro-1,1-dimethylethyl chloroformate (403 mg, 1.68 mmol) andpyridine (0.15 mL, 1.85 mmol) in CH₂Cl₂ (25 mL) followed bytriethylamine (0.43 mL, 3.06 mmol), diphenyl chlorophosphate (0.48 mL,2.30 mmol) and 4-pyrrolidinopyridine (100 mg) to afford 1.37 g (70%) of2-(trimethylsilyl)ethyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-undecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethyethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.0 (s, 9H), 0.88 (m, 8H),1.1-1.7 (m, 44H), 1.80 (s, 3H), 1.89 (s, 3H), 2.23 (m, 6H), 3.58 (m,3H), 4.32 (m, 1H), 4.71 (m, 2H), 4.83 (d, 1H, J=12.1 Hz), 5.01 (d, 1H,J=8.1 Hz), 5.20 (m, 1H), 5.62 (m, 2H), 7.25 (m, 10H).

(4) In the same manner as described in Example 13-(4), the compoundprepared in (4) above (1.28 g, 1.0 mmol) was deprotected with TFA (5 mL)and then treated with the Vilsmeier reagent generated from DMF (0.39 mL,5.0 mmol) and oxalyl chloride (0.22 mL, 2.5 mmol) to give 1.12 g (93%)of2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-undecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosylchloride as a white foam: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.7 Hz),1.1-1.55 (m, 44H), 1.78 (s, 3H), 1.88 (s, 3H), 2.18 (m, 2H), 2.43 (m,2H), 4.34 (m, 4H), 4.72 (m, 3H), 5.09 (m, 1H), 5.50 (t, 1H, J=9.6 Hz),5.80 (d, 1H, J=8.0 Hz), 6.26 (d, 1H, J=3.4 Hz), 7.26 (m, 10H).

(5) In the same manner as described in Example 13-(5), compoundsprepared in (1) and (4) above (530 mg, 0.90 mmol, and 1.0 g, 0.83 mmol,respectively) were coupled in the presence of AgOTf (1.16 g, 4.5 mmol)to afford 1.11 g (76%) ofN-[(R)-3-undecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-undecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-13-D-glucopyranosyl]-L-serinebenzyl ester: ¹H NMR (CDCl₃) δ 0.88 (m, 12H), 1.0-1.65 (m, 88H), 1.77(s, 3H), 1.85 (s, 3H), 2.1-2.5 (m, 8H), 3.37 (m, 1H), 3.64 (m, 1H), 3.85(m, 1H), 4.30 (m, 3H), 4.78 (m, 5H), 5.18 (m, 4H), 5.46 (m, 1H), 6.07(m, 1H), 6.62 (d, 1H, J=7.7 Hz), 7.05-7.45 (m, 15H).

(6) In the same manner as described in Example 2-(7), the compoundprepared in (5) above (1.0 g, 0.57 mmol) was deprotected with zinc (2.0g, 30.5 mmol) and acylated with (R)-3-undecanoyloxytetradecanoic acid(280 mg, 0.68 mmol) in the presence of EEDQ (185 mg, 0.75 mmol) toafford 470 mg (50%) ofN-[(R)-3-undecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-2-[(R)-3-undecanoyloxytetradecanoylamino]-3-O-((R)-3-undecanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinebenzyl ester as an amorphous solid.

(7) In the same manner as described in Example 2-(8), the compoundprepared in (6) above (470 mg, 0.27 mmol) was hydrogenated in thepresence of palladium hydroxide on carbon in EtOH (10 mL) and platinumoxide (400 mg) in EtOH/AcOH (10:1) to afford 130 mg (30%) ofN-[(R)-3-undecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-undecanoyloxytetradecanoylamino)-3-O-[(R)-3-undecanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinetriethylammonium salt as a white powder: mp 181-183° C.; IR (film) 3294,2923, 2853, 1734, 1655, 1466, 1377, 1163, 1080, 721 cm⁻¹; ¹H NMR(CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.8 Hz), 1.1-1.7 (m, 117H), 2.2-2.7 (m,12H), 3.06 (q, 6H, J=7.1 Hz), 3.4-3.2 (m, 5H), 3.6-3.9 (m, 4H), 4.20 (d,1H, 9.8 Hz), 4.54 (d, 1H, J=8.0 Hz), 4.62 (br. S, 1H), 5.17 (m, 4H); ¹³CNMR (CDCl₃) δ 173.5, 173.3, 172.8, 172.2, 169.6, 169.1, 101.5, 77.2,74.8, 0.9, 69.2, 60.5, 58.5, 53.1, 51.5, 46.1, 41.5, 41.1, 39.2, 34.6,34.4, 34.1, 32.0, 29.8, 9.7, 29.4, 29.2, 25.6, 25.2, 25.1, 22.7, 18.5,14.2, 8.7.

Anal. Calcd. for C₉₀H₁₇₂N₃O₁₉P: C, 66.26;H, 10.63; N, 2.58; P, 1.90.Found: C, 66.56;H, 10.57; N, 2.47; P, 1.91.

EXAMPLE 15 B14 PREPARATION OFN-[(R)-3-DECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-D-SERINETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₉H₁₉CO, X═Y═O,N═M═P—Q═O, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

(1) In the same manner as described in Example 2-(5), D-serine benzylester (390 mg, 2.0 mmol) was acylated with(R)-3-decanoyloxytetradecanoic acid (875 mg, 2.2 mmol) in the presenceof EDC·MeI (745 mg, 2.5 mmol) in CH₂Cl₂ to afford 1.05 g (91%) ofN-[(R)-3-decanoyloxytetradecanoyl]-D-serine benzyl ester: mp 51-52° C.;¹H NMR (CDCl₃) δ 0.88 (m, 6H), 1.1-1.7 (m, 34H), 2.30 (t, 2H, J=7.7 Hz),2.50 (m, 2H),3.68 (s, 1H), 3.93 (d, 2H, J=3.1 Hz), 4.62 (m, 1H), 5.22(m, 3H), 6.63 (d, 1H, J=6.9 Hz), 7.35 (br s, 5H).

(2) In the same manner as described in Example 2-(2), the compoundprepared in Example 2-(1) (1.0 g, 2.02 mmol) was acylated with(R)-3-decanoyloxytetradecanoic acid (884 mg, 2.22 mmol) in the presenceof EDC·MeI (720 mg, 2.4 mmol) and 4-pyrrolidinopyridine (100 mg) inCH₂Cl₂, and then deprotected in aqueous AcOH (25 mL) to afford 1.30 g(77%) of 2-(trimethylsilyl)ethyl2-deoxy-3-O-[(R)-3-decanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.00 (s, 9H), 0.88 (m, 8 H),1.25 (m, 30H), 1.59 (m, 4H), 2.30 (t, 2H, J=7.5 Hz), 2.52 (m, 2H), 3.42(m, 1H), 3.55 (m, 1H), 3.66 (m, 1H), 3.83 (dd, 1H, J=11.8, 4.6 Hz), 3.94(m, 2H), 4.57 (d, 1H, J=8.2 Hz), 4.71 (m, 2H), 5.07 (m, 2H), 5.27 (d,1H, J=8.8 Hz).

(3) In the same manner as described in Example 2-(3), the compoundprepared in (2) above (1.25 g, 1.50 mmol) was treated with2,2,2-trichloro-1,1-dimethylethyl chloroformate (396 mg, 1.65 mmol) andpyridine (0.15 mL, 1.81 mmol) in CH₂Cl₂ (25 mL) followed bytriethylamine (0.42 mL, 3.00 mmol), diphenyl chlorophosphate (0.47 mL,2.25 mmol) and 4-pyrrolidinopyridine (100 mg) to afford 1.31 g (69%) of2-(trimethylsilyl)ethyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.0 (s, 9H), 0.89 (m, 8H),1.1-1.7 (m, 34H), 1.82 (s, 3H), 1.90 (s, 3H), 2.30 (m, 4H), 3.40 (q, 1H,J=9.6 Hz), 3.65 (m, 1H), 3.89 (m, 1H), 4.32 (m, 2H), 4.63 (m, 2H), 4.82(d, 1H, J=12.1 Hz), 5.01 (d, 1H, J=8.2 Hz), 5.63 (m, 2H), 7.29 (m, 10H).

(4) In the same manner as described in Example 13-(4), the compoundprepared in (3) above (1.27 g, 1.0 mmol) was deprotected with TFA (5 mL)and then treated with the Vilsmeier reagent generated from DMP (0.39 mL,5.0 mmol) and oxalyl chloride (0.22 mL, 2.5 mmol) to give 1.06 g (89%)of2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosylchloride as a white foam: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.6 Hz),1.1-1.55 (m, 34 H), 1.78 (s, 3H), 1.88 (s, 3H), 2.18 (t, 2H, J=7.7 Hz),2.43 (m, 2H), 4.32 (m, 4H), 4.71 (m, 3H), 4.83 (m, 3H), 5.09 (m, 1H),5.50 (t, 1H, J=9.5 Hz), 5.77 (d, 1H, J=8.0 Hz), 6.26 (d, 1H, J=3.4 Hz),7.20 (m, 10H).

(5) In the same manner as described in Example 13-(5), compoundsprepared in (1) and (4) above above (520 mg, 0.90 mmol, and 1.0 g, 0.84mmol, respectively) were coupled in the presence of AgOTf (1.16 g, 4.5mmol) to afford 1.13 g (78%) ofN-[(R)-3-decanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-lucopyranosyl]-D-serinebenzyl ester: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.6 Hz), 1.1-1.65 (m,68H), 1.82 (s, 3H), 1.89 (s, 3H), 2.2-2.6 (m, 8H), 3.40 (m, 1H), 3.64(m, 1H), 4.01 (m, 2H), 4.27 (m, 2H), 4.44 (d, 1H, J=7.1 Hz), 4.60 (m,2H), 4.77 (in, 2H), 5.19 (m, 6H), 6.61 (d, 1H, J=8.3 Hz), 7.05-7.45 (m,15H).

(6) In the same manner as described in Example 2-(7), the compoundprepared in (5) above (1.0 g, 0.58 mmol) was deprotected with zinc (1.9g, 29 mmol) and acylated with (R)-3-decanoyloxytetradecanoic acid (280mg, 0.70 mmol) in the presence of EEDQ (190 mg, 0.77 mmol) to afford 420mg (44%) ofN-[(R)-3-decanoyloxytetradecanoyl3-O-deoxy-4-O-diphenylphosphono-2-[(R)-3-decanoyloxytetradecanoylainino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-β-D-glucopyranosyl]-D-serinebenzyl ester as an amorphous solid.

(7) In the same manner as described in Example 2-(8), the compoundprepared in (6) above (420 mg, 0.25 mmol) was hydrogenated in thepresence of palladium hydroxide on carbon in EtOH (10 mL) and platinumoxide (400 mg) in EtOH/AcOH (10:1) to afford 118 mg (30%) ofN-[(R)-3-decanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-β-D-glucopyranosyl]-D-serinetriethylammonium salt as a white powder: mp 179-181° C.; IR (film) 3283,3100, 2921, 2852, 1732, 1660, 1651, 1564, 1556, 1464, 1417, 1378, 1322,1181, 1061, 856, 722 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.8Hz), 1.1-1.7 (m, 111H), 2.2-2.7 (m, 12H), 3.06 (m, 6H), 3.33 (m, 5H),3.78 (m, 2H), 3.95 (m, 2H), 4.22 (m, 1H), 4.45 (d, 1H, J=7.5 Hz), 4.68(br. s, 1H), 5.13 (m, 3H), 5.26 (m, 1H); ¹³C NMR (CDCl₃) δ δ 173.7,173.5, 173.1, 171.1, 169.9, 100.3, 75.1, 73.9, 71.9, 71.1, 70.9, 70.2,60.9, 53.9, 52.7, 46.0, 41.3, 40.8, 39.4, 34.6, 34.4, 31.9, 29.8, 29.7,29.5, 29.4, 25.6, 25.4, 25.2, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C₈₇H₁₆₆N₃O₁₉P: C, 65.75;H, 10.53; N, 2.64; P, 1.95.Found: C, 65.32;H, 10.28; N, 2.53; P, 1.89.

EXAMPLE 16 B15 PREPARATION OFN-[(R)-3-DECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT. (COMPOUND (I), R₁═R₂═R₃═N—C₉H₁₉CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

(1) In the same manner as described in Example 2-(5), L-serine benzylester (250 mg, 1.08 mmol) was acylated with(R)-3-decanoyloxytetradecanoic acid (478 mg, 1.2 mmol) in the presenceof EDC·MeI (357 mg, 1.2 mmol) in CH₂Cl₂ to afford 0.52 g (84%) ofN-[(R)-3-heptanoyloxytetradecanoyl]-L-serine benzyl ester: mp 52-53° C.;¹H NMR (CDCl₃) δ 0.87 (t, 6H, J=6.9 Hz), 1.1-1.7 (m, 34H), 2.29 (t, 2 H,J=7.5 Hz), 2.49 (d, 2H, J=5.8 Hz), 3.67 (s, 1H), 3.97 (m, 2H), 4.63 (m,1H), 5.19 (m, 3H), 6.61 (d, 1H, J=7.1 Hz), 7.35 (brs, 5H).

(2) In the same manner as described in Example 13-(5), the compoundprepared in (1) above (500 mg, 0.87 mmol), and the compound prepared inExample 15-(4) (1.08 g, 0.90 mmol) were coupled in the presence of AgOTf(1.16 g, 4.5 mmol) to afford 1.35 g (89%) ofN-[(R)-3-decanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosyl]-L-serinebenzyl ester: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.6 Hz), 1.0-1.65 (m,68H), 1.77 (s, 3H), 1.85 (s, 3H), 2.1-2.5 (m, 8H), 3.38 (q, 1H, J=9.1Hz), 3.65 (m, 1H), 3.84 (m, 1H), 4.27 (m, 3H), 4.70 (m, 5H), 4.84 (m,4H), 5.14 (m, 3H), 5.46 (t, 1H, J=9.7 Hz), 6.07 (m, 1H), 6.62 (d, 1H,J=8.0 Hz), 7.05-7.45 (m, 15H).

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (600 mg, 0.34 mmol) was deprotected with zinc(1.13 g, 17.2 mmol) and acylated with (R)-3-decanoyloxytetradecanoicacid (150 mg, 0.38 mmol) in the presence of EEDQ (124 mg, 0.50 mmol) toafford 362 mg (60%) ofN-[(R)-3-decanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinebenzyl ester as an amorphous solid.

(4) In the same manner as described in Example 2-(8), the compoundprepared in (3) above (300 mg, 0.17 mmol) was hydrogenated in thepresence of palladium on carbon (100 mg) and platinum oxide (200 mg) inTBF/AcOH (10:1) to afford 120 mg (44%) ofN-[(R)-3-decanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinetriethylammonium salt as a white powder: mp 175-176° C.; IR (film) 3304,2956, 2923, 2853, 1733, 1654, 1541, 1466, 1377, 1164, 1107, 1080, 845,721 cm ¹; H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18 H, J=6.9 Hz), 1.1-1.7(m,111H), 2.2-2.75 (m, 12H), 3.07 (q,6H, J=7.2 Hz), 3.37 (m, 1H), 3.5-3.95(m, 8H), 4.21 (q, 1H, 11.0 Hz), 4.54 (d, 1H, J=8.9 Hz), 4.61 (br. s,1H), 5.17 (m, 4H), 7.10 (d, 1H, J=9.0 Hz), 7.43 (d, 1H, J=7.9 Hz); ¹³CNMR (CDCl₃) δ 176.3, 173.4, 173.2, 172.8, 172.0, 169.6, 169.2, 101.4,74.7, 70.9, 69.3, 60.4, 53.2, 51.6, 46.1, 41.4, 41.0, 39.1, 34.5, 34.3,34.2, 34.1, 31.9, 29.8, 29.7, 29.6, 29.4, 29.3, 29.2, 25.5, 25.1, 25.0,22.7, 14.1, 8.6.

Anal. Calcd. for C₈₇H₁₆₆N₃O₁₉P H₂O: C, 65.01;H, 10.54; N, 2.61; P, 1.93.Found: C, 64.92;H, 10.38; N, 2.58; P, 2.06.

EXAMPLE 17 B16 PREPARATION OFN-[(R)-3-NONANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-O-(R)-3-NONANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-NONANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT. (COMPOUND (I), R₁═R₂═R₃═N—C₈H₁₇CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

(1) In the same manner as described in Example 2-(5), L-serine benzylester (390 mg, 2.0 mmol) was acylated with(R)-3-nonanoyloxytetradecanoic acid (780 mg, 2.2 mmol) in the presenceof EDC·MeI (845 mg, 2.5 mmol) in CH₂Cl₂ to afford 1.0 g (89%) ofN-[(R)-3-nonanoyloxytetradecanoyl]-L-serine benzyl ester: mp 52-53° C.;¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.6 Hz), 1.1-1.7 (m, 32H), 2.30 (t, 2 H,J=7.7 Hz), 2.51 (d, 2H, J=5.8 Hz), 2.62 (t, 1H, J=6.0 Hz), 3.98 (m, 2H),4.65 (m, 1H), 5.19 (m, 3H), 6.58 (d, 1H, J=6.8 Hz), 7.35 (br s, 5H).

(2) In the same manner as described in Example 2-(2), the compound 5prepared in Example 2-(1) (1.0 g, 2.02 mmol) was acylated with(R)-3-nonanoyloxytetradecanoic acid (852 mg, 2.22 mmol) in the presenceof EDC·MeI (720 mg, 2.4 mmol) and 4-pyrrolidinopyridine (100 mg) inCH₂Cl₂, and then deprotected in aqueous AcOH (25 mL) to afford 1.31 g(79%) of 2-(trimethylsilyl)ethyl2-deoxy-3-O-[(R)-3-nonanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.00 (s, 9H), 0.88 (m, 8 H),1.25 (m, 28H), 1.59 (m, 4H), 2.30 (t, 2H, J=7.5 Hz), 2.52 (m, 2H), 3.42(m, 1 H), 3.55 (m, 1H), 3.66 (m, 1H), 3.83 (dd, 1H, J=11.8, 4.6 Hz),3.94 (m, 2H), 4.57 (d, 1H, J=8.2 Hz), 4.71 (m, 2H), 5.07 (m, 2H), 5.27(d, 1H, J=8.8 Hz).

(3) In the same manner as described in Example 2-(3), the compoundprepared in (2) above (1.25 g, 1.52 mmol) was treated with2,2,2-trichloro-1,1-dimethylethyl chloroformate (400 mg, 1.67 mmol) andpyridine (0.15 mL, 1.84 mmol) in CH₂Cl₂ (25 mL) followed bytriethylamine (0.42 mL, 3.04 mmol), diphenyl chlorophosphate (0.47 mL,2.28 mmol) and 4-pyrrolidinopyridine (100 mg) to afford 1.30 g (67%) of2-(trimethylsilyl)ethyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-nonanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.0 (s, 9H), 0.88 (m, 8H),1.1-1.7 (m, 32H), 1.82 (s, 3H), 1.89 (s, 3H), 2.22 (m, 6H), 3.33 (m,1H), 3.53 (m, 1H), 3.80 (m, 1H), 3.96 (m, 1H), 4.31 (m, 2H), 4.55 (m,2H), 4.83 (d, 1H, J=12.0 Hz), 5.01 (d, 1H, J=7.9 Hz), 5.62 (m, 1H), 7.28(m, 10H).

(4) In the same manner as described in Example 13-(4), the compoundprepared in (3) above (1.26 g, 1.0 mmol) was deprotected with TFA (5 mL)and then treated with the Vilsmeier reagent generated from DMF (0.39 mL,5.0 mmol) and oxalyl chloride (0.22 mL, 2.5 mmol) to give 1.07 g (91%)of2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-nonanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosylchloride as a white foam: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.9 Hz),1.25-1.55 (m, 32H), 1.78 (s, 3H), 1.88 (s, 3H), 2.18 (t, 2H, J=7.7 Hz),2.43 (m, 2H), 4.34 (m, 4H), 4.70 (m, 3H), 4.83 (m, 3H), 5.09 (m, 1H),5.51 (t, 1H, J=10.2 Hz), 5.78 (d, 1H, J=8.0 Hz), 6.25 (d, 1H, J=3.6 Hz),7.19 (m, 10H).

(5) In the same manner as described in Example 13-(5), compoundsprepared in (1) and (4) above (505 mg, 0.90 mmol, and 1.0 g, 0.85 mmol,respectively) were coupled in the presence of AgOTf (1.16 g, 4.5 mmol)to afford 1.03 g (71%) ofN-[(R)-3-nonanoyloxytetradecanoyl]-O-(2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-nonanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosyl]-L-serinebenzyl ester: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.9 Hz), 1.0-1.65 (m,64H), 1.78 (s, 3H), 1.82 (s, 3H), 2.1-2.5 (m, 8H), 3.38 (m, 1H), 3.64(m, 1H), 3.83 (m, 1H), 4.25 (m, 3H), 4.73 (m, 5 H), 5.18 (m, 5H), 6.07(m, 1H), 6.60 (d, 1H, J=7.8 Hz), 7.05-7.45 (m, 15H).

(6) In the same manner as described in Example 2-(7), the compoundprepared in (5) above (1.0 g, 0.59 mmol) was deprotected with zinc (1.93g, 29.5 mmol) and acylated with (R)-3-nonanoyloxytetradecanoic acid (273mg, 0.71 mmol) in the presence of EEDQ (195 mg, 0.78 mmol) to afford 405mg (42%) ofN-[(R)-3-nonanoyloxytetradecanoyl]-O-[deoxy-4-O-diphenylphosphono-2-[(R)-3-nonanoyloxytetradecanoylamino]-3-O-[(R)-3-nonanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinebenzyl ester as an amorphous solid.

(7) In the same manner as described in Example 2-(8), the compoundprepared in (6) above (405 mg, 0.25 mmol) was hydrogenated in thepresence of palladium hydroxide on carbon in EtOH (10 mL) and platinumoxide (400 mg) in EtOH/AcOH (10:1) to afford 185 mg (48%) ofN-[(R)-3-nonanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-nonanoyloxytetradecanoylaminol-3-O-[(R)-3-nonanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinetriethylammonium salt as a white powder: mp 177-179° C.; IR (film) 3306,2955, 2923, 2853, 1732, 1660, 1538, 1467, 1378, 1252, 1165, 1106, 1080,960, 844, 722 cm⁻¹; ¹H NMR (CDCl₁₃—CD₃OD) δ 0.88 (t, 18H, J=6.8 Hz),1.1-1.7 (m, 105H), 2.2-2.75 (m, 12H), 3.07 (q, 6H, J=7.1 Hz), 3.2-3.5(m, 5H), 3.85 (m, 4H), 4.23 (d, 1H, 10.2 Hz), 4.51 (d, 1H, J=8.0 Hz),4.64 (br. s, 1H), 5.18 (m, 4H); ¹³C NMR (CDCl₃) δ 173.3, 172.8, 172.2,169.6, 169.1, 101.5, 74.8, 70.9, 70.8, 69.3, 60.5, 53.2, 51.5, 46.1,41.5, 41.0, 39.2, 34.5, 34.3, 34.1, 32.0, 31.9, 29.8, 29.6, 29.4, 29.3,25.6, 25.2, 25.1, 22.7, 14.1, 8.7.

Anal. Calcd. for C₈₄H₁₆₀N₃O₁₉P: C, 65.21;H, 10.42; N, 2.72; P, 2.00.Found: C, 65.48;H, 10.32; N, 2.62; P, 2.12.

EXAMPLE 18 B17 PREPARATION OFN-[(R)-3-OCTANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-OCTANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-OCTANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₇H₁₅CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

(1) In the same manner as described in Example 2-(5), L-serine benzylester (390 mg, 2.0 mmol) was acylated with(R)-3-octanoyloxytetradecanoic acid (815 mg, 2.2 mmol) in the presenceof EDC·MeI (745 mg, 2.5 mmol) in CH₂Cl₂ to afford 1.02 g (93%) ofN-[(R)-3-octanoyloxytetradecanoyl]-L-serine benzyl ester: mp 50-51° C.;¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.8 Hz), 1.1-1.7 (m, 30H), 2.30 (t, 2 H,J=7.7 Hz), 2.51 (d, 2H, J=5.8 Hz), 2.60 (t, 1H, J=6.0 Hz), 3.97 (m, 2H),4.65 (m, 1H), 5.22 (m, 3H), 6.61 (d, 1H, J=6.9 Hz), 7.35 (br s, 5H).

(2) In the same manner as described in Example 2-(2), the compoundprepared in Example 2-(1) (1.0 g, 2.02 mmol) was acylated with(R)-3-octanoyloxytetradecanoic acid (821 mg, 2.22 mmol) in the presenceof EDC·MeI (720 mg, 2.4 mmol) and 4-pyrrolidinopyridine (100 mg) inCH₂Cl₂, and then deprotected in 90% aqueous AcOH (25 mL) to afford 1.35g (83%) of 2-(trimethylsilyl)ethyl2-deoxy-3-O-[(R)-3-octanoyloxytetradecanoyl)-2-(2,2,2-tlichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.00 (s, 9H), 0.88 (m, 8 H),1.25 (m, 26H), 1.60 (m, 4H), 2.30 (t, 2H, J=7.5 Hz), 2.53 (m, 2H), 3.42(m, 1H), 3.53 (m, 1H), 3.66 (m, 1H), 3.83 (dd, 1H, J=11.8, 4.4 Hz), 3.94(m, 2H), 4.56 (d, 1H, J=8.3 Hz), 4.64 (d, 1H, J=11.8 Hz), 4.77 (d, 1H,J=11.8 Hz), 5.08 (m, 2 H), 5.30 (br. s, 1H).

(3) In the same manner as described in Example 2-(3), the compoundprepared in (2) above (1.30 g, 1.61 mmol) was treated with2,2,2-trichloro-1,1-dimethylethyl chloroformate (425 mg, 1.77 mmol) andpyridine (0.16 mL, 1.95 mmol) in CH₂Cl₂ (25 mL) followed bytriethylamine (0.45 mL, 3.22 mmol), diphenyl chlorophosphate (0.50 mL,2.42 mmol) and 4-pyrrolidinopyndine (100 mg) to afford 1.42 g (71%) of2-(trimethylsilyl)ethyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-octanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.0 (s, 9H), 0.88 (m, 8H),1.1-1.7 (m, 30H), 1.82 (s, 3H), 1.89 (s, 3H), 2.23 (m, 6H), 3.37 (m,1H), 3.65 (m, 1H), 3.83 (m, 1H), 3.96 (m, 1H), 4.55 (m, 2H), 4.83 (d,1H, J=11.8 Hz), 5.01 (d, 1H, J=8.2 Hz), 5.20 (m, 1H), 7.29 (m, 10H).

(4) In the same manner as described in Example 13-(4), the compoundprepared in (3) above (1.24 g, 1.0 mmol) was deprotected with TFA (5 mL)and then treated with the Vilsmeier reagent generated from DMF (0.39 mL,5.0 mmol) and oxalyl chloride (0.22 mL, 2.5 mmol) to give 1.0 g (87%) of2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-octanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosylchloride as a white foam: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.7 Hz),1.25-1.55 (m, 30H), 1.78 (s, 3H), 1.88 (s, 3H), 2.18 (t, 2H, J=7.7 Hz),2.43 (m, 2H), 4.29 (m, 4 H), 4.72 (m, 3H), 5.09 (m, 1H), 5.51 (t, 1H,J=9.9 Hz), 5.79 (d, 1H, J=7.9 Hz), 6.25 (d, 1H, J=3.5 Hz), 7.29 (m,10H).

(5) In the same manner as described in Example 13-(5), compoundsprepared in (1) and (4) above (490 mg, 0.90 mmol, and 1.0 g, 0.86 mmol,respectively) were coupled in the presence of AgOTf (1.16 g, 4.5 mmol)to afford 0.99 g (69%) ofN-[(R)-3-octanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-octanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosyl)-L-serinebenzyl ester: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.9 Hz), 1.0-1.65 (m,60H), 1.77 (s, 3H), 1.85 (s, 3H), 2.1-2.5 (m, 8H), 3.37 (m, 1H), 3.65(m, 1H), 3.83 (m, 1H), 4.27 (m, 3H), 4.72 (m, 5H), 5.18 (m, 4H), 5.46(t, 1H, J=9.8 Hz), 6.06 (m, 1H), 6.60 (d, 1H, J=8.0 Hz), 7.05-7.45 (m,15H).

(6) In the same manner as described in Example 2-(7), the compoundprepared in (5) above (0.95 g, 0.57 mmol) was deprotected with zinc(1.86 g, 28.5 mmol) and acylated with (R)-3-octanoyloxytetradecanoicacid (252 mg, 0.68 mmol) in the presence of EEDQ (185 mg, 0.75 mmol) toafford 433 mg (47%) ofN-[(R)-3-octanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-2-[(R)-3-octanoyloxytetradecanoylarnino]-3-O-[(R)-3-octanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinebenzyl ester as an amorphous solid.

(7) In the same manner as described in Example 2-(8), the compoundprepared in (6) above (433 mg, 0.27 mmol) was hydrogenated in thepresence of palladium hydroxide on carbon (250 mg) in EtOH (10 mL) andplatinum oxide (400 mg) in EtOH/AcOH (10:1) to afford 196 mg (48%) ofN-[(R)-3-octanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-octanoyloxytetradecanoylamino]-3-O-t(R)-3-octanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinetriethylammonium salt as a white powder: mp 177-178° C.; IR (film) 3296,2956, 2923, 2853, 1732, 1645, 1546, 1466, 1378, 1315, 1170, 1082, 1056,961, 846, 722 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.6 Hz),1.1-1.7 20 (m, 99H), 2.2-2.75 (m, 12H), 3.08 (q, 6H, J=7.1 Hz), 3.39 (d,1H, J=8.8 Hz), 3.6 -4.0 (m, 8H), 4.22 (q, 1H, 10.3 Hz), 4.53 (d, 1H,J=8.2 Hz), 4.63 (m, 1H), 5.18 (m, 4H), 7.04 (d, 1H, J=8.8 Hz), 7.42 (d,1H, J=8.0 Hz); ¹³C NMR (CDCl₃) δ 176.8, 173.3, 173.2, 172.7, 172.2,169.6, 169.1, 101.5, 74.8, 70.9, 70.8, 69.3, 60.5, 53.2, 51.5, 46.2,41.5, 41.1, 39.2, 34.5, 34.3, 34.1, 34.0, 32.0, 31.8, 29.8, 29.6, 29.4,29.3, 29.2, 29.1, 25.6, 25.3, 25.2, 25.0, 22.7, 14.1, 8.7.

Anal. Calcd. for C₈₁H₁₅₄N₃O₁₉P.H₂O: C, 63.87;H, 10.32; N, 2.76; P, 2.03.Found: C, 63.96;H, 10.29; N, 2.69; P, 1.67.

EXAMPLE 19 B18 PREPARATION OFN-[(R)-3-HEPTANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-HEPTANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-HEPTANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₆H₁₃CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

(1) In the same manner as described in Example 2-(5), L-serine benzylester (390 mg, 2.0 mmol) was acylated with(R)-3-heptanoyloxytetradecanoic acid (780 mg, 2.2 mmol) in the presenceof EDC·MeI (745 mg, 2.5 mmol) in CH₂Cl₂ to afford 0.97 g (91%) ofN-[(R)-3-heptanoyloxytetradecanoyl]-L-serine benzyl ester: mp 46-48° C.;¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.9 Hz), 1.1-1.7 (m, 28H), 2.30 (t, 2 H,J=7.7 Hz), 2.50 (d, 2H, J=5.8 Hz), 2.62 (t, 1H, J=6.0 Hz), 3.97 (m, 2H),4.65 (m, 1H), 5.19 (m, 3H), 6.61 (d, 1H, J=6.9 Hz), 7.35 (br s, 5H).

(2) In the same manner as described in Example 2-(2), the compoundprepared in Example 2-(1) (1.0 g, 2.02 mmol) was acylated with(R)-3-heptanoyloxytetradecanoic acid (790 mg, 2.22 mmol) in the presenceof EDC·MeI (720 mg, 2.4 mmol) and 4-pyrrolidinopyridine (100 mg) inCH₂Cl₂, and then deprotected in 90% aqueous AcOH (25 mL) to afford 1.30g (81%) of 2-(trimethylsilyl)ethyl2-deoxy-3-O-[(R)-3-heptanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamno)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.00 (s, 9H), 0.88 (m, 8 H),1.25 (m, 24H), 1.59 (m, 4H), 2.30 (t, 2H, J=7.5 Hz), 2.52 (m, 2H), 3.42(m, 1 H), 3.55 (m, 1H), 3.66 (m, 1H), 3.83 (dd, 1H, J=11.5, 4.2 Hz),3.94 (m, 2H), 4.57 (d, 1H, J=8.3 Hz), 4.64 (d, 1H, J=12.1 Hz), 4.76 (d,1H, J=11.9 Hz), 5.09 (m, 2 H), 5.31 (d, 1H, J=8.7 Hz).

(3) In the same manner as described in Example 2-(3), the compoundprepared in (2) above (1.25 g, 1.58 mmol) was treated with2,2,2-trichloro-1,1-dimethylethyl chloroformate (417 mg, 1.74 mmol) andpyridine (0.15 mL, 1.91 mmol) in CH₂Cl₂ (25 mL) followed bytriethylamine (0.44 mL, 3.16 mmol), diphenyl chlorophosphate (0.49 mL,2.37 mmol) and 4-pyrrolidinopyridine (100 mg) to afford 1.34 g (69%) of2-(trimethylsilyl)ethyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-heptanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.0 (S, 9H), 0.88 (m, 8H),1.1-1.7 (m, 28H), 1.82 (s, 3H), 1.89 (s, 3 H), 2.35 (m, 4H), 3.37 (m,1H), 3.61 (m, 1H), 3.80 (m, 1H), 4.32 (m, 2H), 4.63 (m, 2 H), 4.83 (d,1H, J=12.0 Hz), 5.01 (d, 1H, J=8.2 Hz), 5.62 (m, 2H), 7.29 (m, 10H).

(4) In the same manner as described in Example 13-(4), the compoundprepared in (3) above (1.23 g, 1.0 mmol) was deprotected with TFA (5 mL)and then treated with the Vilsmejer reagent generated from DNF (0.39 mL,5.0 mmol) and oxalyl chloride (0.22 mL, 2.5 mmol) to give 1.0 g (87%) of2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-heptanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosylchloride as a white foam: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.9 Hz),1.25-1.55 (m, 28H), 1.78 (s, 3H), 1.88 (s, 3H), 2.18 (t, 2H, J=7.6 Hz),2.43 (m, 2H), 4.26 (m, 4 15H), 4.73 (m, 3H), 5.09 (m, 1H), 5.51 (t, 1H,J=10.2 Hz), 5.77 (d, 1H, J=8.0 Hz), 6.25 (d, 1H, J=3.3 Hz), 7.19 (m,10H).

(5) In the same manner as described in Example 13-(5), compoundsprepared in (1) and (4) above (480 mg, 0.90 mmol, and 0.98 g, 0.86 mmol,respectively) were coupled in the presence of AgOTf (1.16 g, 4.5 mmol)to afford 1.06 g (75%) ofN-[(R)-3-heptanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-heptanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosyl]-L-serinebenzyl ester: ¹H NMR (CDCl₃) δ 0.88 (m, 12H), 1.0-1.65 (m, 56H), 1.77(s, 3H), 1.85 (s, 3H), 2.1-2.5 (m, 8H), 3.38 (m, 1H), 3.64 (m, 1H), 3.83(m, 1H), 4.25 (m, 3H), 4.78 (m, 5H), 5.16 (m, 4H), 5.46 (t, 1H, J=9.9Hz), 6.06 (m, 1H), 6.60 (d, 1H, J=7.7 Hz), 7.05-7.45 (m, 15H).

(6) In the same manner as described in Example 2-(7), the compoundprepared in (5) above (1.0 g, 0.61 mmol) was deprotected with zinc (2.0g, 30.5 mmol) and acylated with (R)-3-heptanoyloxytetradecanoic acid(260 mg, 0.73 mmol) in the presence of EEDQ (200 mg, 0.80 mmol) toafford 440 mg (45%) ofN-[(R)-3-heptanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-2-[(R)-3-heptanoyioxytetradecanoylamino]-3-O-[(R)-3-heptanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinebenzyl ester as an amorphous solid.

(7) In the same manner as described in Example 2-(8), the compoundprepared in (6) above (440 mg, 0.28 mmol) was hydrogenated in thepresence of palladium hydroxide on carbon (250 mg) in EtOH (10 mL) andplatinum oxide (400 mg) in EtOH/AcOH (10:1) to afford 208 mg (51%) ofN-[(R)-3-heptanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-heptanoyloxytetradecanoylamino]-3-O-[(R)-3-heptanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinetriethylammonium salt as a white powder: mp 176-177° C.; IR (film) 3307,2956, 2924, 2854, 1732, 1650, 1545, 1466, 1378, 1316, 1170, 1080, 956,841, 722 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (m, 18H), 1.1-1.7 (m, 93H),2.2-2.75 (m, 12H), 3.08 (q, 6H, J=7.2 Hz), 3.40 (d, 1H, J=10.2 Hz),3.6-4.0 (m, 7 H), 4.24 (m, 2H), 4.52 (d, 1H, J=8.0 Hz), 4.63 (m, 1H),5.19 (m, 4H), 7.04 (d, 1H, J=8.6 Hz), 7.40 (d, 1H, J=8.4 Hz); ¹³C NMR(CDCl₃) δ 177.1, 173.2, 173.1, 172.7, 172.3, 169.5, 168.9, 101.5, 75.074.8, 71.2, 70.9, 69.1, 60.5, 53.1, 51.4, 46.1, 41.5, 41.0, 39.2, 34.5,34.3, 34.1, 34.0, 31.9, 31.6, 31.5, 29.8, 29.6, 29.4, 29.0, 28.9, 28.8,25.6, 25.3, 25.1, 25.0, 22.7, 22.6, 14.1, 8.7.

Anal. Calcd. for C₇₈H₁₄₈N₃O₁₉P: C, 64.04;H, 10.20; N, 2.87; P, 2.12.Found: C, 63.77;H, 10.1 1; N, 2.85; P, 2.02.

EXAMPLE 20 B19 PREPARATION OF2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]ETHYL2-DEOXY-4-O-PHOSPHONO-3-O-[(R)-3-TETRADECANOYOXYTETRADECANOYL]-2-[(R)-3-TETRADECANOYOXYTETRADECANOYLAMINO]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₃H₂₇CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₆═R₇═R₉═H, R₈═PO₃H₂).

(1) 2-Amino-1-(t-butyldiphenylsilyloxy)ethane (330 mg, 1.1 mmol) and(R)-3-tetradecanoyloxytetradecanoic acid (500 mg, 1.1 mmol) weredissolved in CH₂Cl₂ (10 mL) and treated with powdered 4 A molecularsieves (500 mg). After 1 h EEDQ (297 mg, 1.2 mmol) was added and thereaction was stirred for 18 h, filtered through Celite® and concentratedin vacuo. The residue was chromatographed over silica gel using 15%EtOAc/hexanes to give 675 mg (92%) of a colorless solid. A portion ofthis material (500 mg, 0.68 mmol) was deprotected with TBAF (1 M in THF,1 mL, 1 mmol) in THF (5 mL) by stirring at room temperature for 2 h. Thereaction mixture was diluted with Et₂O (50 mL) and washed with brine(2×50 mL). The brine was back extracted with Et₂O (2×50 mL) and thecombined organic extracts were dried over Na₂SO₄ and concentrated invacuo to afford 338 mg (62%) of2-[(R)-3-tetradecanoloxytetradecanoylamino]ethanol as an off-whitesolid.

(2) In the same manner as described in Example 2-(6), the compoundprepared in (1) above (338 mg, 0.68 mmol) and the compound prepared inExample 2-(4) (786 mg, 0.61 mmol) were coupled in the presence ofmercury cyanide (770 mg, 3.05 mmol) to give 245 mg (24%) of2-[(R)-3-tetradecanoyloxytetradecanoylamino]ethyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.9 Hz), 1.1-1.8(m, 84H), 1.81 (s, 3H), 1.89 (s, 3 H), 2.15-2.55 (m, 8H), 3.25 (m, 1H),3.47 (m, 2H), 3.67 (mn, 1H), 3.83 (m, 2H), 4.28 (dd, 1H, J=12.2, 4.9Hz), 4.36 (d, 1H, J=11.0 Hz), 4.68 (m, 2H), 4.78 (d, 1H, J=11.6Hz),4.94(d, 1H, J=11.6 Hz),5.16(m,2H), 5.53 (t, 1H, J=10.0 Hz),6.06 (d,1H, J=4.9 Hz), 6.19 (m, 1H), 7.25 (m, 10H).

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (500 mg, 0.29 mmol) was deprotected with zinc (980mg, 15 mmol) and then acylated with (R)-3-tetradecanoyloxytetradecanoicacid (155 mg, 0.34 mmol) in the presence of EEDQ (110 mg, 0.44 mmol) togive 315 mg (62%) of 2-[(R)-3-tetradecanoyloxytetradecanoylamino]ethyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-2-[(R)-3-tetradecanoyoxytetradecanoylamino]-β-D-glucopyranosideas an amorphous solid.

(4) In the same manner as described in Example 2-(8), the compoundprepared in (3) above (200 mg, 0.113 mmol) was hydrogenated in thepresence of platinum oxide (100 mg) to give 142 mg (76%) of2-[(R)-3-tetradecanoyloxytetradecanoylamino]ethyl2-deoxy-4-O-phosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-2-[(R)-3-tetradecanoyoxytetradecanoylamino]-β-D-glucopyranosidetriethylammonium salt as a white solid: mp 175-176° C.; IR (film) 3285,3098, 2955, 2919, 2851, 1731, 1659, 1642, 1556, 1468, 1379, 1250, 1228,1174, 1110, 1083, 1046, 962, 857 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t,18H, J=6.0 Hz), 1.1-1.7 (m, 135H), 2.2-2.7 (m, 15H), 3.06 (q, 6H, J=7.1Hz), 3.2-4.1 (m, 8 H), 4.21 (q, 1H, J=9.9 Hz), 4.51 (d, 1H, J=8.2 Hz),5.05-5.25 (m, 4H), 7.33 (d, 1H, J=8.5 Hz), 7.50 (br t, 1H, J=4.8 Hz);¹³C NMR (CDCl₃) δ 173.7, 173.3, 170.6, 170.3, 169.9, 100.9, 75.8, 73.0,71.3, 71.1, 70.9, 70.6, 68.3, 60.6, 55.1, 45.7, 41.6, 41.2, 39.5, 34.6,34.5, 34.4, 32.0, 29.8, 29.4, 29.3, 25.4, 25.1, 22.7, 14.2, 8.6.

Anal. Calcd. for C₉₈H₁₉(N₃O₁₇P.2H₂O: C, 67.28;H, 11.18; N, 2.40; P,1.77. Found: C, 67.01;H, 11.18; N, 2.15; P, 2.01.

EXAMPLE 21 B20 PREPARATION OF2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]ETHYL2-DEOXY-4-O-PHOSPHONO-3-O-[(R)-3-DECANOYOXYTETRADECANOYL]-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₉H₁₉CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₆═R₇═R₉═H, R₈═PO₃H₂).

(1) In the same manner as described in Example 20-(1),2-amino-1-(t-butyldiphenylsilyloxy)ethane (450 mg, 1.5 mmol) wasacylated with (R)-3-decanoyloxytetradecanoic acid (600 mg, 1.5 mmol) inthe presence of EDC·MeI (594 mg, 2.0 mmol) and then deprotected withTBAF (1.0 M in THF, 2.5 mL, 2.5 mmol) in THF (10 mL) to afford 488 mg(81%) of 2-[(R)-3-decanoyloxytetradecanoylamino]ethanol as an off-whitesolid.

(2) In the same manner as described in Example 13-(5), the compoundprepared in (1) above (385 g, 0.87 mmol) and the compound prepared inExample 15-(4) (1.05 g, 0.87 mmol) were coupled in the presence of AgOTf(560 mg, 2.2 mmol) to give 1.04 g (74%) of2-[(R)-3-decanoyloxytetradecanoylamino]ethyl2-deoxy-4-O-diphenylphosphono-3-O-t(R)-3-decanoyoxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.9 Hz), 1.1-1.6(m, 68H), 1.78 (s, 3H), 1.88 (s, 3H), 2.18 (t, 2H, J=7.7 Hz), 2.44 (m,2H), 4.34 (m, 5H), 4.72 (m, 2H), 4.83 (q, 1H, J=9.3 Hz), 5.09 (m, 1H),5.51 (t, 1H, J=10.2 Hz), 5.79 (d, 1H, J=8.0 Hz), 6.26 (d, 1H, J=3.4 Hz),7.31 (m, 10H).

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (700 mg, 0.44 mmol) was deprotected with zinc(1.42 g, 21.7 mmol) and then acylated with(R)-3-decanoyloxytetradecanoic acid (190 mg, 0.48 mmol) in the presenceof EEDQ (148 mg, 0.6 mmol) to give 432 mg (62%) of2-1(R)-3-decanoyloxytetradecanoylamino]ethyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyoxytetradecanoyl]-2-[(R)-3-decanoyloxytetradecanoylamino]-β-D-glucopyranosideas an amorphous solid.

(4) In the same manner as described in Example 2-(8), the compoundprepared in (3) above (400 mg, 0.25 mmol) was hydrogenated in thepresence of platinum oxide (200 mg) to give 200 mg (52%) of2-[(R)-3-decanoyloxytetradecanoylamino]ethyl2-deoxy-4-O-phosphono-3-O-((R)-3-decanoyoxytetradecanoyl]-2-[(R)-3-decanoyloxytetradecanoylamino]-β-D-glucopyranosidetriethylammonium salt as a white solid: mp 165-166° C.; IR (film) 3289,3094, 2956, 2922, 2853, 1732, 1658, 1644, 1556, 1467, 1379, 1247, 1164,1107, 1081, 1048 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.9 Hz),1.1-1.7 (m, 111H), 2.2-2.7 (m, 15H), 3.05 (q, 6H, J=7.1 Hz), 3.2-3.85(m, 9H), 4.52 (d, 1H, J=8.2 Hz), 5.05-5.25 (m, 4H), 7.21 (d, 1H, J=8.5Hz), 7.42 (br t, 1H); ¹³C NMR (CDCl₃) δ 173.8, 173.3, 170.7, 170.3,170.0, 100.9, 75.6, 73.0, 71.3, 70.9, 70.6, 68.3, 60.7, 55.0, 45.8,41.6, 41.2, 39.5, 34.5, 34.4, 34.1, 31.9, 29.8, 29.6, 29.5, 29.4, 25.4,25.1, 22.7, 14.2, 8.6.

Anal. Calcd. for C₈₆H₁₆₆N₃O₁₇P.H₂O: C, 66.08;H, 10.83; N, 2.69; P, 1.98.Found: C, 65.80;H, 10.63; N, 2.63; P, 2.04.

EXAMPLE 22 B21 PREPARATION OF3-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]PROPYL2-DEOXY-4-O-PHOSPHONO-3-O-[(R)-3-TETRADECANOYOXYTETRADECANOYL]-2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₃H₂₇CO, X═Y═O, N═1,M═P═Q═O, R₄═R₅═R₆═R₇═R₉═H, R₈═PO₃H₂).

(1) In the same manner as described in Example 20-(1),3-amino-1-(t-butyldiphenylsilyloxy)propane (470 mg, 1.5 mmol) wasacylated with (R)-3-tetradecanoyloxytetradecanoic acid (680 mg, 1.5mmol) in the presence of EDC·MeI (595 mg, 2.0 mmol) and then deprotectedwith TBAF (1.0 M in THF, 2.0 mL, 2.0 mmol) in THF (10 mL) to afford 698mg (91%) of 3-[(R)-3-tetradecanoyloxytetradecanoylamino]-1-propanol asan off-white solid.

(2) In the same manner as described in Example 13-(4), the compoundprepared in Example 2-(3) (7.9 g, 5.88 mmol) was deprotected with TFA(10 mL) and then treated with the Vilsmeier reagent generated from DMF(1.8 mL, 23.5 mmol) and oxalyl chloride (1.03 mL, 11.76 mmol) in CH₂Cl₂(60 mL) to give 6.32 g (85%) of2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosylchloride as a white foam: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.8 Hz),1.2-1.55 (m, 42H), 1.78 (s, 3H), 1.88 (s, 31H), 2.18 (t, 2H, J=7.5 Hz),2.43 (m, 211), 4.31 (m, 41H), 4.68 (d, 1H, J=11.9 Hz), 4.74 (d, 1H,J=11.9 Hz), 4.83 (q, 1H, J=9.3 Hz), 5.09 (m, 1H), 5.51 (t, 1H, J=9.7Hz), 5.78 (d, 1H, J=8.0 Hz), 6.26 (d, 1H, J=3.4 Hz), 7.31 (m, 10H).

(3) In the same manner as described in Example 13-(5), the compoundprepared in (1) above (613 mg, 1.2 mmol) and the compound prepared in(2) above (1.5 g, 1.2 mmol) were coupled in the presence of AgOTf (642mg, 2.5 mmol) to give 1.43 g (68%) of3-[(R)-3-tetradecanoyloxytetradecanoylaminolpropyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.9 Hz), 1.1-1.8(m, 86H), 1.82 (s, 3H), 1.89 (s, 3H), 2.20 (t, 2H, J=7.6 Hz), 2.29 (t,2H, J=7.7 Hz), 2.44 (m, 4H), 3.21 (m, 1H), 3.42 (m, 1H), 3.54 (m, 2H),3.80 (m, 1H), 3.94 (m, 1H), 4.28 (dd, 1H, J=12.3, 5.2 Hz), 4.38 (d, 1H,J=10.8 Hz), 4.70 (m, 3H), 4.81 (d, 1H, J=8.2 Hz), 5.14 (m, 2H), 5.47 (t,1H, J=9.6 Hz), 6.13 (d, 1H, J=7.6 Hz), 6.22 (br. s, 1H), 7.25 (m, 10H).

(4) In the same manner as described in Example 2-(7), the compoundprepared in (3) above (700 mg, 0.40 mmol) was deprotected with zinc(1.32 g, 20.1 mmol) and then acylated with(R)-3-tetradecanoyloxytetradecanoic acid (200 mg, 0.44 mmol) in thepresence of EEDQ (125 mg, 0.5 mmol) to give 435 mg (60%) of3-[(R)-3-tetradecanoyloxytetradecanoylamino]propyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-2-[(R)-3-tetradecanoyloxytetradecanoylamino])-β-D-glucopyranoside as an amorphous solid.

(5) In the same manner as described in Example 2-(8), the compoundprepared in (4) above (400 mg, 0.22 mmol) was hydrogenated in thepresence of platinum oxide (200 mg) to give 170 mg (45%) of3-[(R)-3-tetradecanoyloxytetradecanoylaminolpropyl2-deoxy-4-O-phosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-2-[(R)-3-tetradecanoyloxytetradecanoylamino])-β-D-glucopyranosidetriethylammonium salt as a white solid: mp 171-172° C.; IR (film) 3288,3094, 2955, 2919, 2850, 1731, 1658, 1344, 1556, 1468, 1378, 1320, 1251,1226, 1172, 1106, 1083, 1044 cm⁻¹; LH NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H,J=6.0 Hz), 1.1-1.7 (m, 135H), 2.2-2.7 (m, 15H), 3.06 (q, 6H, J=7.1 Hz),3.2-4.1 (m, 8H), 4.21 (q, 1H, J=9.9 Hz), 4.51 (d, 1 H. J=8.3 Hz),5.05-5.25 (m, 4H), 7.23 (t, 1H, J=5.3 Hz), 7.33 (d, 1H, J=8.6 Hz); ¹³CNMR (CDCl₃) δ 173.5, 173.4, 170.6, 170.2, 169.9, 100.6, 75.8, 71.5,70.9, 70.5, 66.8, 60.4, 55.3, 45.6, 41.4, 39.4, 36.3, 34.6, 34.5, 34.2,31.9, 29.7, 29.4, 29.3, 29.1, 25.4, 25.1, 22.7, 14.1, 8.5.

Anal. Calcd. for C₉₉H₁₉₂N₃O₁₇P.2H₂O: C, 67.42;H, 11.20; N, 2.38; P,1.76. Found: C, 66.97;H, 11.01; N, 2.38; P, 1.95.

EXAMPLE 23 B22 PREPARATION OF4-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]BUTYL2-DEOXY-4-O-PHOSPHONO-3-O-[(R)-3-TETRADECANOYOXYTETRADECANOYL]-2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO])-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₃H₂₇CO, X═Y═O, N=2,M═P═Q═O, R₄═R₅═R₆═R₇═R₉═H, R₈═PO₃H₂).

(1) In the same manner as described in Example 20-(1),4-amino-1-(t-butyldiphenylsilyloxy)butane (500 mg, 1.53 mmol) wasacylated with (R)-3-tetradecanoyloxytetradecanoic acid (695 mg, 1.53mmol) in the presence of EDC·MeI (595 mg, 2.0 mmol) and then deprotectedwith TBAF (1.0 M in THF, 2.5 mL, 2.5 mmol) in THF (15 mL) to afford 651mg (81%) of 4-[(R)-3-tetradecanoyloxytetradecanoylamino]-1-butanol as anoff-white solid.

(2) In the same manner as described in Example 13-(5), the compoundprepared in (1) above (650 mg, 1.25 mmol) and the compound prepared inExample 22-(2) (1.6 g, 1.25 mmol) were coupled in the presence of AgOTf(1.16 g, 4.5 mmol) to give 1.65 g (75%) of4-[(R)-3-tetradecanoyloxytetradecanoylamino]butyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.9 Hz), 1.1-1.8(m, 88H), 1.82 (s, 3H), 1.89 (s, 3H), 2.15-2.55 (m, 8H), 3.24 (m, 2H),3.50 (m, 2H), 3.83 (m, 2H), 4.27 (dd, 1H, J=12.1, 3.8 Hz), 4.32 (d, 1H,J=11.5 Hz), 4.66 (m, 2H), 4.78 (d, 1H, J=12.1 Hz), 4.89 (d, 1H, J=8.0Hz), 5.15 (m, 2H), 5.54 (t, 1H, J=9.7 Hz), 5.95 (m, 2H), 7.25 (m, 10H).

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (700 mg, 0.39 mmol) was deprotected with zinc(1.30 g, 19.8 mmol) and then acylated with(R)-3-tetradecanoyloxytetradecanoic acid (195 mg, 0.43 mmol) in thepresence of EEDQ (125 mg, 0.5 mmol) to give 421 mg (60%) of4-[(R)-3-tetradecanoyloxytetradecanoylamino]butyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-2-[(R)-3-tetradecanoyloxytetradecanoylarino])-β-D-glucopyranosideas an amorphous solid.

(4) In the same manner as described in Example 2-(8), the compoundprepared in (3) above (400 mg, 0.22 mmol) was hydrogenated in thepresence of platinum oxide (200 mg) to give 212 mg (55%) of4-[(R)-3-tetradecanoyloxytetradecanoylamino]butyl2-deoxy-4-O-phosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-2-[(R)-3-tetradecanoyloxytetradecanoylamino])-β-D-glucopyranosidetriethylammonium salt as a white solid: mp 171-172° C.; IR (film) 3298,2955, 2920, 2851, 1732, 1645, 1550, 1467, 1378, 1181, 1107, 1083, 1044,721 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.9 Hz), 1.1-1.7 (m,135H), 2.2-2.7 (m, 19H), 3.05 (q, 6H, J=7.1 Hz), 3.18 (m, 2H), 3.3-3.5(m, 6H), 3.78 (m, 3 H), 3.97 (d, 1H, J=12.5 Hz), 4.23 (q, 1H, J=10.0Hz), 4.50 (d, 1H, J=8.5 Hz), 5.13 (m, 4H), 7.12 (d, 1H, J=9.1 Hz); ¹³CNMR (CDCl₃) δ 173.9, 173.4, 173.3, 170.8, 169.9, 169.8, 101.0, 75.6,73.2, 71.4, 71.1, 70.6, 68.9, 60.7, 54.8, 45.9, 41.5, 39.6, 38.9, 34.6,34.3, 32.0, 29.8, 29.5, 29.0, 28.9, 26.3, 25.4, 25.1, 22.7, 14.2, 8.7.

Anal. Calcd. for C₁₀₀H₁₉₄N₃O₁₇P.H₂O: C, 68.26;H, 11.23; N, 2.39; P,1.76. Found: C, 68.21;H, 11.03; N, 2.26; P, 1.73.

EXAMPLE 24 B23 PREPARATION OF4-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]HEXYL2-DEOXY-4-O-PHOSPHONO-3-O-[(R)-3-TETRADECANOYOXYTETRADECANOYL]-2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]-β-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₃H₂₇CO, X═Y═O, N=4,M═P═Q═O, R₄═R₅═R₆═R₇═R₉═H, R₈═PO₃H₂).

(1) In the same manner as described in Example 20-(1),6-amino-1-(t-butyldiphenylsilyloxy)hexane (1.48 g, 4.15 mmol) wasacylated with (R)-3-tetradecanoyloxytetradecanoic acid (2.07 g, 4.56mmol) in the presence of EDC·MeI (1.35 g, 4.56 mmol) and thendeprotected with TBAF (1.0 M in THF, 1.53 mL, 1.53 mmol) in THF (46 mL)to afford 700 mg (30%) of6-[(R)-3-tetradecanoyloxytetradecanoylamino]-1-hexanol as an off-whitesolid.

(2) In the same manner as described in Example 13-(5), the compoundprepared in (1) above (689 mg, 1.20 mmol) and the compound prepared inExample 22-(2) (1.25 g, 1.00 mmol) were coupled in the presence of AgOTf(1.28 g, 5.0 mmol) to give 1.59 g (94%) of4-[(R)-3-tetradecanoyloxytetradecanoylamino]hexyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDC₃) δ 0.88 (t, 12H, J=6.6 Hz), 1.1-1.8(m, 92H), 1.82 (s, 3H), 1.89 (s, 3H), 2.22 (t, 2H, J=7.6 Hz), 2.29 (t,2H, J=7.4 Hz), 2.45 (m, 4H), 3.22 (m, 1H), 3.46 (m, 2H), 3.83 (m, 2H),3.94 (m, 1H), 4.31 (m, 2H), 4.64 (m, 2H), 4.83 (d, 1H, J=12.1 Hz), 4.97(d, 1H, J=7.8 Hz), 5.17 (m, 2H), 5.59 (t, 1H, J=8.8 Hz), 5.75 (m, 1H),5.84 (d, 1H, J=7.6 Hz), 7.25 (m, 10H).

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (1.57 g, 0.88 mmol) was deprotected with zinc(2.88 g, 44.1 mmol) and then acylated with(R)-3-tetradecanoyloxytetradecanoic acid (481 mg, 1.06 mmol) in thepresence of EEDQ (327 mg, 1.32 mmol) to give 1.57 g (97%) of4-[(R)-3-tetradecanoyloxytetradecanoylamino]hexyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-2-[(R)-3-tetradecanoyloxytetradecanoylamino])-β-D-glucopyranosideas an amorphous solid.

(4) In the same manner as described in Example 2-(8), the compoundprepared in (3) above (1.57 g, 0.85 mmol) was hydrogenated in thepresence of platinum oxide (157 mg) to give 130 mg (10%) of4-[(R)-3-tetradecanoyloxytetradecanoylamino]hexyl2-deoxy-4-O-phosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-2-[(R)-3-tetradecanoyloxytetradecanoylamino]-β-D-glucopyranosidetriethylammonium salt as a white solid: mp 150-152° C.; IR (film) 3284,3099, 2954, 2920, 2851, 1731, 1657, 1637, 1557, 1467, 1418, 1378, 1320,1249, 1179, 1108, 1083, 1044, 856, 721 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.89(t, 18H, J=6.6 Hz), 1.1-1.7 (m, 135H), 2.2-2.7 (m, 23H), 3.05 (q, 6H,J=7.1 Hz), 3.18 (m, 2 H), 3.39 (d, 1H, J=8.2 Hz), 3.49 (q, 1H, J=7.5Hz), 3.82 (m, 2H), 3.99 (d, 1H, J=11.9 Hz), 4.25 (q, 1H, J=8.9 Hz), 4.59(m, 2H), 5.18 (m, 4H); ¹³C NMR (CDCl₃) δ 173.7, 173.3, 170.6, 169.7,169.4, 100.6, 75.5, 73.1, 71.3, 70.9, 70.6, 69.2, 60.6, 55.2, 45.8,41.7, 41.4, 39.5, 39.4, 34.6, 34.3, 34.2, 34.1, 31.9, 29.7, 29.4, 29.2,26.5, 25.5, 25.3, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C₁₀₂H₁₉₈N₃O₁₇P.H₂O: C, 68.53;H, 11.28; N, 2.33; P,1.73. Found: C, 68.63 H, 11.12; N, 2.26; P, 1.66.

EXAMPLE 25 B24 PREPARATION OFN-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-O-PHOSPHONO-2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-L-SERINAMIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₃H₂₇CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CONH₂, R₈═PO₃H₂).

(1) A suspension of L-serinamide hydrochloride (0.157 g, 1.18 mmol) and(R)-3-tetradecanoyloxytetradecanoic acid (0.61 g, 1.34 mmol) in CH₂Cl₂(6 mL) was treated with triethylamine (0.18 mL, 1.3 mmol) and theresulting solution was stirred with 4 Å molecular sieves for 30 min.EEDQ (0.437 g, 1.77 mmol) was then added and the mixture was stirred for16 h at room temperature. The product that precipitated was collectedand washed with CH₂Cl₂ (2×25 mL) to give 0.455 g (71%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-L-serinamide as a colorlesspowder: mp 126-130° C.; ¹H NR (CDCl₃) δ 0.88 (t, 6H, J=7 Hz), 1.15-1.7(m, 42H), 2.31 (t, 2H, J=7.5 Hz), 2.51 (d, 2H, J=6.3 Hz), 3.56 (br s,1H), 3.65 (dd, 1H, J=11.2, 5.5 Hz), 3.86 (dd, 1H, J=11.2, 4.5 Hz), 4.21(s, 2H), 4.40 (m, 1H), 5.22 (m, 1H).

(2) In the same manner as described in Example 2-(6), the compoundprepared in (1) above (0.23 g, 0.246 mmol) and the compound prepared inExample 2-(4) (0.961 g, 0.745 mmol) were coupled in the presence ofmercury cyanide (0.43 g, 1.7 mmol) to give 0.527 g (71%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2,-trichloroethoxycarbonylamino)-β-D-glucopyranosyl]-L-serinamideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=7H), 1.0-1.7 (m,84H), 1.80 and 1.89 (2s, 6H), 2.21 (t, 2H, J=7.5 Hz), 2.30 (t, 2H, J=7.5Hz), 2.37 (m, 2H), 2.47 (m, 2H), 3.54 (m, 1H), 3.68 (dd, 1H, J=8, J=11Hz), 3.86 (br d, 1H, J=11 Hz), 4.16 (dd, 1H, J=11, 4 Hz), 4.24 (dd, 1H,J=12, 4.3 Hz), 4.40 (d, 1H, J=12 Hz), 4.6-4.8 (m, 4H), 5.00 (d, 1H, J=8Hz), 5.1-5.25 (m, 2H), 5.4-5.55 (m, 2H), 5.84 (br s, 1H), 6.61 (br s,2H), 7.1-7.35 (m, 10H).

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (0.44 g, 0.254 mmol) was deprotected with zinc(0.83 g, 13 mmol) and then acylated with(R)-3-tetradecanoyloxytetradecanoic acid (0.14 g, 0.31 mmol) in thepresence of EEDQ (0.095 g, 0.38 mmol) to give 0.271 g (59%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-2-[(R)-3-tetradecanoyloxytetradecanoylamino]-3-O-t(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinamideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 18 H, J=6.5 Hz),1.0-1.7 (m, 126H), 2.03 (br s, 1H), 2.15-2.55 (m, 12H), 3.5-4.05 (m,5H), 4.14 (dd, 1H, J=10, 3.5 Hz), 4.5-4.65 (m, 2H), 4.68 (d, 1H, J=8.1Hz), 5.05-5.25 (m, 3H), 5.31 (t, 1H, J=10 Hz), 5.58 (br s, 1H), 6.31 (d,1H, J=8 Hz), 6.85-6.95 (m, 2H), 7.1-7.4 (m, 10H).

(4) In the same manner as described in Example 2-(8), the compoundprepared in (3) above (0.25 g, 0.14 mmol) was hydrogenated in thepresence of platinum oxide (0.125 g) to give 0.195 (80%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-tetradecanoyloxytetradecanoylaminol-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinamidetriethylammonium salt as a colorless solid: mp 190-191° C. (dec); IR(film) 3418, 3293, 2921, 2850, 1732, 1717, 1651, 1636, 1557, 1540, 1458,1165, 1033 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=7 Hz), 1.0-1.7(m, 135H), 2.2-2.7 (m, 12H), 3.05 (q, 6H, J=7.2 Hz), 3.2-3.45 (m),3.5-4.15 (m, 5H), 4.21 (q, 1H, J=10 Hz), 4.53 (d, 1H, J=8.1 Hz), 4.58(m, 1H), 5.0-5.3 (m, 4H), 7.25 (d, 1H, J=8.4 Hz), 7.40 (d, 1H, J=7.2Hz); ¹³C NMR (CDCl₃—CD₃OD) δ 173.7, 173.5, 72.5, 170.7, 170.5, 170.4,101.4, 75.5, 73.4, 71.1, 70.9, 70.2, 68.6, 60.0, 53.9, 52.2, 5.6, 41.2,41.0, 38.9, 34.4, 34.2, 31.8, 29.6, 29.5, 29.3, 29.1, 25.2, 24.9, 22.6,14.0, 8.3.

Anal. Calcd for C₉₉H₁₉₁N₄O₁₈P.2.5H₂O: C, 66.00;H, 10.97; N, 3.11; P, 72.Found: C, 66.04;H, 10.99; N, 3.03; P, 1.95.

EXAMPLE 26 B25 PREPARATION OFN-[(R)-3-DECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-L-SERINAMIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₉H₁₉CO, X═Y═O,N═M═P═Q=O, R₄═R₅═R₇═R₉═H, R₆═CONH₂, R₈═PO₃H₂).

(1) In the same manner as described in Example 25-(1), L-serinamidehydrochloride (169 mg, 1.2 mmol) was acylated with(R)-3-decanoyloxytetradecanoic acid (478 mg, 1.2 mmol) in the presenceof EEDQ (371 mg, 1.5 mmol) in CH₂Cl₂ to afford 428 mg (74%) ofN-[(R)-3-decanoyloxytetradecanoyl]-L-serinamide as a white solid: ¹H NMR(CDCl₃) δ 0.88 (t, 6H), 1.1-1.7 (m, 34H), 2.33 (t, 2H, J=7.5 Hz), 2.54(d, 2H, J=6.6 Hz), 3.35 (s, 2H), 3.72 (dd, 1H, J=11.0, 5.2 Hz), 3.84(dd, 11H, J=11.3, 5.0 Hz), 4.20 (t, 1H, J=5.1 Hz), 5.26 (t, 1H, J=6.4Hz).

(2) In the same manner as described in Example 13-(5), the compoundprepared in (1) above (410 mg, 0.85 mmol) and the compound prepared inExample 15-(4) (1.05 g, 0.87 mmol) were coupled in the presence of AgOTf(560 mg, 2.2 mmol) to afford 780 g (56%) ofN-[(R)-3-decanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosyl]-L-serinamideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H), 1.1-1.6 (m, 68H),1.80 (s, 3H), 1.89 (s, 3H), 2.30 (m, 8H), 3.53 (m, 1H), 3.68 (m, 1H),3.85 (br. d, 1H, J=9.4 Hz), 4.15 (dd, 1H, J=10.8, 3.7 Hz), 4.24 (dd, 1H,J 12.3, 4.6 Hz), 4.40 (d, 1H, J=10.8), 4.65 (m, 4H), 5.00 (d, 1H, J=8.2Hz), 5.18 (m, 2H), 5.46 (m, 2H), 5.83 (m, 1H), 6.60 (m, 2H), 7.30 (m,10H).

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (600 mg, 0.36 mmol) was deprotected with zinc(1.19 g, 18.2 mmol) and acylated with (R)-3-decanoyloxytetradecanoicacid (160 mg, 0.4 mmol) in the presence of EEDQ (124 mg, 0.50 mmol) toafford 371 mg (62%) ofN-[(R)-3-decanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-β-D-glucopyranosyl]-serinamide as an amorphous solid.

(4) In the same manner as described in Example 2-(8), the compoundprepared in (3) above (330 mg, 0.20 mmol) was hydrogenated in thepresence of platinum oxide (200 mg) to afford 120 mg (44%) ofN-[(R)-3-decanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinamidetriethylammonium salt as a white powder: mp 187-189° C; IR (film) 3419,3286, 3220, 3098, 2955, 2922, 2852, 1732, 1680, 1662, 1644, 1559, 1467,1247, 1167, 1107, 1080, 1051, 965, 913 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.89(t, 18H, J=7.0 Hz), 1.1-1.7 (m, 111H), 2.2-2.7 (m, 12H), 3.07 (q, 6H,J=7.1 Hz), 3.68 (m, 1H), 3.87 (m, 1H), 4.09 (dd, 1H, J=10.8, 3.6 Hz),4.22 (m, 1H), 4.53 (d, 1H, J=8.2 Hz), 4.58 (m, 1H), 5.13 (m, 3H), 5.28(m, 1H), 7.53 (d, 1H, J=9.0 Hz), 7.56 (d, 1H, J=7.7 Hz); ¹³C NMR (CDCl₃)δ 173.5, 173.2, 170.2, 169.8, 102.3, 75.7, 73.5, 71.3, 70.7, 70.1, 68.8,60.8, 53.9, 51.7, 45.8, 41.5, 41.1, 39.1, 34.6, 34.5, 34.2, 32.0, 29.7,29.6, 29.5, 29.4, 25.7, 25.4, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C₈₇H₁₆₇N₄O₁₈P.H₂O: C, 65.05;H, 10.60; N, 3.49; P, 1.93.Found: C, 65.06;H, 10.40; N, 3.31; P, 2.00.

EXAMPLE 27 B26 PREPARATION OFN-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-TRADECANOYLOXYTETRADECANOYL]-β-D-GLUCOPYRANOSYL]-1-SERINEMETHYL ESTER TRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₃H₂₇CO,X═Y═O, N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂ME, R₈═PO₃H₂).

(1) A solution of the compound prepared in Example 12-(2) (0.290 g,0.157 mmol) in THF (20 mL) was hydrogenated in the presence of 5%palladium on carbon (50 mg) at room temperature and atmospheric pressurefor 3 h. The catalyst was removed by filtration and the filtrateconcentrated. A solution of the residue in CHCl₃ (5 mL) at 0° C. wastreated with a solution of diazomethane (0.5 mmol) in ether (5 mL) andthen stirred for 30 min at 0° C. AcOH (0.5 mL) was added and theresulting colorless solution was diluted with ether (50 mL), washed withsaturated aqueous NaHCO₃ (25 mL), dried (Na₂SO₄) and concentrated. Flashchromatography on silica gel (gradient elution, 20→25% EtOAcs-hexanes)afforded 0.199 g (72%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichoro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosyl]-L-serinemethyl ester as an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.5Hz), 1.1-1.75 (m, 84H), 1.81 and 1.89 (2s, 6H), 2.36 (t, 2H, J=7.5 Hz),2.25-2.6 (mn, 6H), 3.48 (q, 1H, J=8 Hz), 3.7-3.9 (mn, 5H), 4.2-4.4 (m,3H), 4.6-4.85 (m, 4H), 4.88 (d, 1H, J=7.8 Hz), 5.03-5.22 (m, 2H), 5.49(t, 1H, J=9.5 Hz), 6.21 (br s, 1H), 6.59 (d, 1H, J=7.8 Hz), 7.1-7.4 (mn,10H).

(2) In the same manner as described in Example 2-(7), the compoundprepared in (1) above (0.195 g, 0.111 mmol) was deprotected with zinc(0.36 g, 5.5 mmol) and acylated with (R)-3-tetradecanoyloxytetradecanoicacid (0.060 g, 0.13 mmol) in the presence of EEDQ (0.041 g, 0.17 mmol)to give 0.138 g (69%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-O-[(R)-4-O-diphenylphosphono-2-[(R)-3-tetradecanoyloxytetradecanoylano]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl-β-D-glucopyranosyl]-L-serinemethyl ester as an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 t, 18H, J=6.5Hz), 1.0-1.75 (m, 126H), 2.15-2.45 (m, 10H), 2.52 (dd, 1H, J=14.7, 6Hz), 2.66 (dd, 1H, J=14.7, 6 Hz), 3.35 (br s, 1H), 3.4-3.8 (m, 7H), 3.88(dd, 1H, J=11 Hz), 4.18 (dd, 1H, J=11 Hz), 4.6-4.75 (m, 2H), 5.03 (d,1H, J=7.8 Hz), 5.1-5.25 (m, 3H), 5.50 (t, 1H, J=˜9.5 Hz), 6.50 (d, 1H,J=7.2 Hz), 6.97 (d, 1H, J=7.8 Hz), 7.1-7.4 (m, 10H).

(3) In the same manner as described in Example 2-(8), the compoundprepared in (2) above (0.100 g, 0.055 mmol) was hydrogenated in thepresence of platinum oxide (50 mg) to give 0.055 g (57%) ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-tetradecanoyloxytetradecanoylamino]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinemethyl ester triethylammonium salt as a colorless solid: mp 142-143° C.(dec); IR (film) 3289, 2955, 2921, 2852, 1733, 1718, 1699, 1652, 1558,1540, 1521, 1506, 1469, 1457, 1375, 1360, 1259 cm⁻¹; ¹H NMR(CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=˜6.5 Hz), 1.0-1.7 (m, 135H), 2.2-2.7 (m,12H), 3.05 (q, 6H, J=7.5 Hz), 3.31 (d, 1H, J=9.3 Hz), 3.37 (s, 1H),3.55-3.9 (m, 10H), 3.97 (d, 1H, J=12 Hz), 4.1-4.25 (m, 2H), 4.55-4.65(m, 2H), 5.05-5.25 (m, 3H), 7.23 (d, 1H, J=8.1 Hz), 7.47 (d, 1H, J=7.2Hz); ¹³C NMR (CDCl₃) δ 173.6, 173.4, 170.5, 170.4, 170.1, 100.7, 75.9,72.8, 71.2, 70.8, 70.6, 68.5, 60.3, 55.3, 52.7, 52.4, 47.7, 41.5, 40.9,39.7, 34.6, 34.5, 34.3. 32.0, 29.8, 29.4, 25.4, 25.1, 22.7, 14.2, 8.5.

Anal. Calcd for C₁₀₀H₁₉₂N₃O₁₉P.H₂O: C, 67.11;H, 10.93; N, 2.35; P, 1.73.Found: C, 66.91;H, 10.93; N, 2.31; P, 2.11.

EXAMPLE 28 B27 PREPARATION OFN-(CARBOXYMETHYL)-N-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-2-AMINOETHYL2-DEOXY-4-O-PHOPHONO-2-[(R)-3-TETRADECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-β-d-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₁₃H₂₇CO, X═Y═O,N=M=P=0, R₄═R₅═R₆═R₉═H, R₇═CO₂H, Q=1, R₈═PO₃H₂).

(1) In the same manner as described in Example 2-(5),N-(2-hydroxyethyl)glycine t-butyl ester (0.25 g, 1.43 mmol) was acylatedwith (R)-3-tetradecanoyloxytetradecanoic acid (0.714 g, 1.57 mmol) inthe presence of EDC·MeI (0.466 g, 1.57 mmol) to give 0.46 g (51%) ofN-(2-hydroxyethyl)-N-[(R)-3-tetradecanoyloxytetradecanoyl]glycinet-butyl ester as an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 6H, J=6.5Hz), 1.15-1.7 (m, 51H), 2.26 (t, 2H, J=7.5 Hz), 2.60 (dd, 1H, J=6.5, 15Hz), 2.86 (dd, 1H, J=6.7, 15 Hz), 3.40-4.15 (m, 7H), 5.25 (m, 1H).

(2) In the same manner as described in 13-(5), the compound prepared in(1) above (0.21 g, 0.334 mmol) and the compound prepared in Example22-(2) (0.458 g, 0.368 mmol) were coupled in the presence of AgOTf(0.688 g, 2.68 mmol) to give 0.39 g (64%) ofN-(t-butyloxycarbonylmethyl)-N-[(R)-3-tetradecanoyloxytetradecanoyl]-2-aminoethyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosideas an amorphous solid: ¹H NMR (CDCl₃) δ 0.88 (t, 12H, J=6.5 Hz),1.0-1.95 (m, 99 H), 2.1-2.6 (m, 7H), 2.84 (dd, 1H, J=5, 15 Hz), 3.2-4.15(m, 8H), 4.15-4.45 (m, 2H), 4.55-4.9 (m, 3H), 5.00 (d, 1H, J=8 Hz), 5.13(m, 2H), 5.4-5.65 (m, 1H), 6.16 (d, 1H, J=7 Hz), 7.05-7.4 (m, 10H).

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (0.339 g, 0.185 mmol) was deprotected with zinc(0.36 g, 5.54 mmol) and then acylated with(R)-3-tetradecanoyloxytetradecanoic acid (0.100 g, 0.221 mmol) in thepresence of EEDQ (0.068 g, 0.276 mmol) to give 0.25 g (71%) ofN-(t-butyloxycarbonylmethyl)-N-[(R)-3-tetradecanoyloxytetradecanoyl]-2-aminoethyl2-deoxy-4-O-phosphono-2-[(R)-3-tetradecanoyloxytetradecanoylamino]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosideas a colorless solid.

(4) In the same manner as described in Example 2-(8), the compoundprepared in (3) above (0.25 g, 0.131 mmol) was hydrogenated in thepresence of platinum oxide (125 mg) in 9:1 THEF-AcOH (15 mL). The crudehydrogenolysis product was dissolved in CH₂Cl₂ (1 mL), cooled to 0° C.,and treated dropwise with TFA (0.5 mL). After stirring for 2 h at 0° C.,the reaction mixture was concentrated and residual TFA was removed byazeotroping with toluene. The resulting residue (0.23 g) was dissolvedin 1% aqueous triethylamine (12 mL) and lyophilized. Flashchromatography on silica gel withchloroform-methanol-water-triethylamine (91:8:0.5:0.5→85:15:0.5:0.5,gradient elution) and further purification by means of acidic extractionas described in Example 2-(8) and lyophilization from 1% aqueoustriethylamine (6 mL) afforded 99 mg (43%) ofN-(carboxymethyl)-N-[(R)-3-tetradecanoyloxytetradecanoyl]-2-aminoethyl2-deoxy-4-O-phosphono-2-[(R)-3-tetradecanoyloxytetradecanoylaminol-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as colorless solid: mp 162-163° C. (dec); IR(film) 3286, 2922, 2852, 1732, 1651, 1556, 1455, 1434, 1378, 1260, 1088,801 cm⁻¹; ¹H NMR (CDCl₃) δ 0.88 (t, 18H, J=6.5 Hz), 1.0-1.75 (m, 135H),2.2-3.0 (m, 14H), 3.04 (q, 6H, J=7.2 Hz), 3.25-3.8 (m, 5H), 3.85-4.3 (m,5H), 4.55 (d, 1H, J=7.5 Hz), 4.68 (d, 1H, J=8.1 Hz), 5.05-5.35 (m, 4H).

Anal. Calcd for C₁₀₀H₁₉₂N₃O₁₉P.3H₂O: C, 65.79;H, 10.60; N, 2.30; P,1.70. Found: C, 65.82;H, 10.44; N, 2.40; P, 1.79.

EXAMPLE 29 B28 PREPARATION OFN-CARBOXYMETHYL-N-[(R)-3-DECANOYLOXYTETRADECANOYL]-3-AMINOPROPYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO])-3-O-[(R)-3-DECANOYOXYTETRADECANOYL]-β-d-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₉H₁₉CO, X═Y═O, N=1,M=P=0, R₄═R₅═R₆═R₉═H, R₇═CO₂H, Q=1, R₈═PO₃H₂).

(1) In the same manner as described in Example 2-(5),N-(3-hydroxypropyl)glycine benzyl ester (450 mg, 2.0 mmol) was acylatedwith (R)-3-decanoyloxytetradecanoic acid (1.0 g, 2.5 mmol) in thepresence of EDC·MeI (900 mg, 3.0 mmol) in CH₂Cl₂ to afford 0.76 g (63%)of N-(3-hydroxypropyl)-N-[(R)-3-decanoyloxytetradecanoyl]glycine benzylester as a colorless oil: ¹H NMR (CDCl₃) (1:1 mixture of rotomers) δ0.88 (t, 6H, J=6.6 Hz), 1.1-1.7 (m, 35H), 1.78 (m, 1H), 2.26 (q, 2H,J=7.6 Hz), 2.37 and 2.54 (2 dd, 1H, J=14.9, 6.9 Hz), 2.60 and 2.89 (2dd, 1H, J=14.8, 6.0 Hz), 3.51 (m, 4H), 3.70 (m, 1H), 3.95-4.25 (m, 2H),5.1-5.25 (m, 3H),7.35(m, 5H).

(2) In the same manner as described in Example 13-(5), the compoundprepared in (1) above (500 mg, 0.83 mmol), and the compound prepared inExample 15-(4) (1.0 g, 0.83 mmol) were coupled in the presence of AgOTf(1.07 g, 4.15 mmol) to afford 1.27 g (72%) ofN-(benzyloxycarbonylmethyl)-N-[(R)-3-decanoyloxytetradecanoyl]-3-aminopropyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyoxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-β-D-glucopyranosidebenzyl ester: ¹H NMR (CDCl₃) (2:1 mixture of rotomers) δ 0.88 (t, 12H,J=6.9 Hz), 1.1-1.7 (m, 69H), 1.80 (s, 3H), 1.88 (s, 3H), 2.1-2.6 (m,11H), 2.81 (dd, 1H, J=14.8, 6.2 Hz), 3.37 (m, 1H), 3.52 (m, 2H), 3.76(m, 1H), 3.87 (m, 1H), 4.05 (m, 2H), 4.28 (m, 3H), 4.62 (m, 3H), 4.77(m, 1H), 4.93 (d, 1H, J=8.2 Hz), 5.15 (m, 4H), 5.46 and 5.61 (2 t, 1H,J=9.5 Hz), 5.95 and 6.05 (2 d, 1H, J=7.5 Hz), 7.1-7.4 (m, 15H).

(3) In the same manner as described in Example 2-(7), the compoundprepared in (2) above (1.25 g, 0.71 mmol) was deprotected with zinc(2.31 g, 3.53 mmol) and acylated with (R)-3-decanoyloxytetradecanoicacid (353 mg, 0.89 mmol) in the presence of EEDQ (264 mg, 1.07 mmol) toafford 670 mg (54%) ofN-benzyloxycarbonylmethyl-N-[(R)-3-decanoyloxytetradecanoyl]-3-aminopropyl2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyoxytetradecanoyl]-2-[(R)-3-decanoyloxytetradecanoylamino])-β-D-glucopyranosideas an amorphous solid.

(4) In the same manner as described in Example 2-(8), the compoundprepared in (3) above (670 mg, 0.38 mmol) was hydrogenated in thepresence of palladium hydroxide on carbon (270 mg) and platinum oxide(200 mg) in EtOH/AcOH (10:1) to afford 240 mg (39%) ofN-carboxymethyl-N-[(R)-3-decanoyloxytetradecanoyl]-3-aminopropyl2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylamino])-3-O-[(R)-3-decanoyoxytetradecanoyl]-β-D-glucopyranosidetriethylammonium salt as a white powder: mp 156-157° C.; IR (film) 3284,2929, 2853, 2729, 1732, 1655, 1628, 1551, 1466, 1378, 1314, 1164, 1108,1047, 955, 844, 722 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.9Hz), 1.1-1.7 (m, 111H), 2.27 (q, 6H, J=6.2 Hz), 2.35-2.80 (m, 9H), 3.05(q, 6H, J=7.2 Hz), 3.25-3.60 (m, 4H), 3.75-4.10 (m, 4H), 4.23 (m, 2H),4.47 (d, 1H, J=8.2 Hz), 4.61 (d, 1H, J=8.3 Hz), 5.05-5.25 (m, 4H); ¹³CNMR (CDCl₃) δ 173.4, 173.0, 171.1, 170.6, 170.3, 169.6, 100.5, 74.5,73.9, 71.4, 71.2, 70.7, 70.2, 67.0, 65.8, 60.7, 54.6, 54.3, 51.4, 49.2,46.0, 45.4, 42.1, 41.2, 39.4, 38.0, 37.7, 34.5, 34.3, 34.2, 31.9, 29.8,29.7, 29.6, 29.5, 29.2, 28.1, 25.4, 25.3, 25.1, 22.7, 14.1, 11.1, 8.6.

Anal. Calcd. for C₈₉H₁₇₀N₃O₁₉P.H₂O: C, 65.37;H, 10.60; N, 2.57; P, 1.89.Found: C, 65.35;H, 10.42; N, 2.43; P, 2.05.

EXAMPLE 30 B29 PREPARATION OFN-[(R)-3-HEXANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-HEXANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-HEXANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYL]-L-SERINAMIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₅H₁₁CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₈═CONH₂, R₈═PO₃H₂).

In the same manner as described in Example 26 and cognate steps,N-[(R)-3-hexanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-hexanoyloxytetradecanoylamino]-3-O-[(R)-3-hexanoyloxytetradecanoyl]-β-D-glucopyranosyl]-L-serinamidetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranoside,L-serinamide hydrochloride, and (R)-3-hexanoyloxytetradecanoic acid: mp184-185° C.; IR (film) 3416, 3284, 3210, 3096, 2954, 2923, 2853, 1735,1721, 1680, 1664, 1646, 1560, 1466, 1246, 1169, 1080, 1038 cm⁻¹; ¹H NMR(CDCl₃—CD₃OD) δ 0.90 (m, 18H), 1.1-1.7 (mH), 2.23-2.47 (m, 6H), 2.48-2.7(m, 6H), 3.06 (q, 6H, J=6 Hz), 3.26-3.34 (mH), 3.66 (m, 1H), 3.77 (d,1H, J=9.5 Hz), 3.82-3.96 (m, 2H), 4.12 (dd, 1H, J=2, 8 Hz), 4.21 (q, 1H,J=8 Hz), 4.56 (d, 1H, J=7 Hz), 4.61 (m, 1H), 5.05-5.18 (m, 3H), 5.24 (m,1H), 7.26 (d, 1H, J=6.5 Hz), 7.40 (d, 1H, J=5.7 Hz).

Anal. Calcd. for C₇₅H₁₄₃N₄O₁₈P.H₂O: C, 62.65 H, 10.16; N, 3,90; P, 2.15.Found: C, 62.60 H, 9.97; N, 3.72; P, 2.25.

EXAMPLE 31 B30 PREPARATION OFN-[(R)-3-DECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-HEXANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYL]-L-SERINAMIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₃═N—C₉H₁₉CO R₂═N—C₅H₁₁CO,X═Y═O, N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CONH₂, R₈═PO₃H₂).

In the same manner as described in Example 26 and cognate steps, N-[(R)-3-decanoyloxytetradecanoyl)-O-[2-deoxy-4-O-phosphono-2-[(R)-3-hexanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-(-D-glucopyranosyl]-L-serinamidetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylarnino)-α-D-glucopyranoside,L-serinamide hydrochloride, and (R)-3-hexa- and decanoyloxytetradecanoicacids: mp 200-201° C. dec; IR (film) 3420, 3286,2956,2923,2853,1733,1680,1663,1645, 1556,1466, 1418,1378, 1248,1168, 1106, 1081, 1051859, 722 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.9 Hz), 1.0-1.7(m, 103H), 2.15-2.71 (m, 12H), 3.06 (q, 6H, J=7 Hz), 3.68 (m, 1H), 3.87(m, 1H), 4.09 (dd, 1H, J=10.8, 3.6 Hz), 3.35-4.0 (mH), 4.15-4.3 (m, 2H),4.57-4.7 (m, 2H), 5.05-5.3 (m, 4H), 7.42 (m, 1H); ¹³C NMR (CDCl₃) δ173.5, 173.1, 170.2, 169.8, 102.2, 75.8, 73.7, 71.3, 70.7, 70.2, 69.0,60.7, 53.9, 51.7, 45.8, 41.3, 41.1, 39.1, 34.6, 34.5, 34.2, 32.0, 29.7,32.0, 31.4, 29.8, 29.6, 29.5, 29.4, 25.6, 25.4, 25.1, 24.7, 22.7, 22.4,13.9, 8.6.

Anal. Calcd. for C₈₃H₁₅₉N₄O₁₉P.H₂O: C, 64.31;H, 10.47; N, 3.61. Found:C, 64.31;H, 10.27; N, 3.41.

EXAMPLE 32 B31 PREPARATION OF2-[(R)-3-HEXANOYLOXYTETRADECANOYLAMINO]ETHYL2-DEOXY-4-O-PHOSPHONO-3-O-[(R)-3-HEXANOYLOXYTETRADECANOYL]-2-[(R)-3-HEXANOYLOXYTETRADECANOYLAMINO]-α-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₅H₁₁CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₆═R₇═R₉═H, R₈═PO₃H₂).

In the same manner as described in Example 21 and cognate steps,2-[(R)-3-hexanoyloxytetradecanoylamino]ethyl2-deoxy-4-O-phosphono-3-O-[(R)-3-hexanoyloxytetradecanoyl]-2-[(R)-3-hexanoyloxytetradecanoylamino]-α-D-glucopyranosidetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranoside,2-amino-1-(t-butyldiphenylsilyloxy)ethane, and(R)-3-hexanoyloxytetradecanoic acid: mp 161-162° C.; IR (film) 3288,3096, 2956, 2924, 2854, 1732, 1657, 1645, 1557, 1466, 1378, 1316, 1245,1173, 1080, 1041 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.89 (m, 18H), 1.15-1.67(m, 87H), 2.23-2.70 (m, 15H), 3.06 (q, 6H, J=7.4 Hz), 3.2-3.85 (n, 9H),4.52 (d, 1H, J=8.0 Hz), 5.05-5.27 (m, 4H), 7.24 (d, 1H, J=8.5 Hz), 7.43(br t, 1H); ¹³C NMR (CDCl₃) δ 173.7, 173.3, 173.3, 170.6, 170.2, 169.9,100.9, 75.6, 73.0, 71.3, 70.9, 70.6, 68.3, 60.7, 55.0, 45.8, 41.6, 41.2,39.5, 34.5, 34.4, 34.4, 31.9, 31.3, 29.7, 29.4, 25.4, 24.7, 22.7, 22.4,14.1, 8.6.

Anal. Calcd. for C₇₄H₁₄₂N₃O₁₇P.H₂O: C, 63.72;H, 10.40; N, 3.01; P, 2.22.Pound: C, 63.72;H, 10.21; N, 2.96; P, 2.46.

EXAMPLE 33 B32 PREPARATION OF2-[(R)-3-HEXADECANOYLOXYTETRADECANOYLAMINO]ETHYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-OCTADECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═C₁₃H₂₇CO, R₂═C₁₇H₃₅CO,R₃═N—C₁₅H₃₁CO, X═Y═O, N=M=P=Q=0, R₄═R₅═R₆═R₇═R₉═H, R₈═PO₃H₂).

In the same manner as described in Example 21 and cognate steps,2-[(R)-3-hexadecanoyloxytetradecanoylamino]ethyl2-deoxy-4-O-phosphono-2-[(R)-3-octadecanoyloxytetradecanoylanino]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-α-D-glucopyranosidetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranoside,2-amino-1-(t-butyldiphenylsilyloxy)ethane, and (R)-3-tetra-, octa- andhexadecanoyloxytetradecanoic acids: mp 180-181° C.; IR (film) 3284,3097, 2920, 2851, 1731, 1657, 1699, 1683, 1653, 1558, 1541, 1521, 1506,1467, 1435, 1418, 1376, 1258, 1173, 1033 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ0.8-1.75 (mH), 2.2-2.7 (mH), 3.08 (q, 6H, J=7.2 Hz), 3.2-3.5 (m, 5H),3.55-4.05 (mH), 4.24 (q, 1H, J=7 Hz), 4.53 (d, 1H, J=8 Hz), 5.05-5.3 (m,4H), 7.32 (d, 1H, J=9 Hz), 7.49 (br t, 1H); ¹³C NMR (CDCl₃) δ 173.8,173.4, 173.3, 170.6, 170.3, 169.9, 100.9, 75.7, 73.0, 71.3, 70.9, 70.6,68.3, 60.7, 55.0, 45.8, 41.3, 39.5, 34.6, 34.4, 32.0, 29.8, 29.4, 25.4,25.1, 22.7, 14.2,8.6.

Anal. Calcd. for C₁₀₄H₂₀₂N₃O₁₇P.4H₂O: C, 66.81;H, 11.32; N, 2.25. Found:C, 66.52;H, 10.80; N, 2.19.

EXAMPLE 34 B34 PREPARATION OFN-[(R)-3-HEXANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-HEXANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-HEXANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYL]-L-SERINETRIETHLAMMONIUM SALT. (COMPOUND (I), R₁═R₂═R₃═N—C₅H₁₁CO, X═Y═O,N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

In the same manner as described in Example 16 and cognate steps, N-[(R)-3-hexanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-hexanoyloxytetradecanoylamino]-3-O-[(R)-3-hexanoyloxytetradecanoyl]-α-D-glucopyranosyl]-L-serinetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranoside,L-serine benzyl ester, and (R)-3-hexanoyloxytetradecanoic acid: mp159-160° C.; IR (film) 3317, 2954, 2924, 2854, 1734, 1654, 1540, 1466,1377, 1316, 1245, 1173, 1082, 846, 722 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88(m, 18H), 1.15-1.7 (mH), 2.2-2.75 (m, 12H), 3.08 (q, 6H, J=7.2 Hz), 3.40(d, 1H, J=9.9 Hz), 3.55-3.95 (mH), 4.15-4.3 (m, 1H), 4.51 (d, 1H, J=8.0Hz), 4.63 (br. s, 1H), 5.1-5.3 (m, 4H), 7.01 (d, 1H, J=9.1 Hz), 7.37 (d,1 H, J=8.8 Hz); 13C NMR (CDCl₃) δ 177.0, 173.2, 173.2, 172.7, 172.3,169.6, 169.0, 101.5, 75.0, 71.2, 70.9, 70.8, 69.1, 60.5, 53.1, 51.4,46.1, 41.4, 41.0, 39.1, 34.5, 34.2, 34.1, 34.0, 31.9, 31.4, 31.3, 29.8,29.6, 29.4, 25.6, 25.3, 25.1, 24.7, 24.7, 22.7, 22.5, 22.4, 14.1, 14.0,8.7.

Anal. Calcd. for C₇₅H₁₄₂N₃O₁₉P.2H₂O: C, 61.83;H, 10.10; N, 2.88; P,2.13. Found: C, 62.07;H, 10.01; N, 2.94; P, 2.40.

EXAMPLE 35 B35 PREPARATION OFN-[(R)-3-DECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]3-O-[(R)-3-HEXANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYL]-L-SERINETRIETYLAMMONIUM SALT. (COMPOUND (I), R₁═N—C₅H₁₁CO, R₂═R₃═N—C₉H₁₉CO,X═Y═O, N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₆═PO₃H₂).

In the same manner as described in Example 16 and cognate steps,N-[(R)-3-decanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-hexanoyloxytetradecanoyl]-α-D-glucopyranosyl]-L-serinetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranoside,L-serine benzyl ester, and (R)-3-hexa- and decanoyloxytetradecanoicacids: mp 158-159° C.; IR (film) 3304, 2956, 2923, 2853, 1732, 1658,1547, 1466, 1378, 1317, 1246, 1174, 1082, 960, 846, 722 cm⁻¹; ¹H NMR(CDCl₃—CD₃OD) δ 0.88 (m, 18H), 1.15-1.7 (mH), 2.2-2.75 (m, 12H), 3.06(q, 6H, J=7.2 Hz), 3.3-3.63 (mH), 3.66-3.98 (m, 4H), 4.1-4.3 (m, 2H),4.54 (d, 1H, J=8.0 Hz), 4.6 (m, 1H), 5.05-5.27 (m, 4H), 7.15 (d, 1H,J=8.7 Hz), 7.46 (d, 1H, J=8.2 Hz); ¹³C NMR (CDCl₃) δ 173.6, 173.3,172.8, 172.1, 169.6, 169.2, 101.5, 74.8, 70.9, 70.8, 69.3, 60.5, 53.2,51.5, 46.1, 41.9, 41.5, 41.0, 39.2, 34.5, 34.3, 34.1, 31.9, 31.4, 29.8,29.6, 29.4, 25.6, 25.3, 25.1, 25.1, 25.0, 24.8, 22.7, 22.5, 14.1, 11.1,8.7.

Anal. Calcd. for C₈₃H₁₅₈N₃O₁₉P.H₂O: C, 64.27;H, 10.40; N, 2.71; P, 2.00.Found: C, 64.14;H, 10.33; N, 2.70; P, 2.05.

EXAMPLE 36 B36 PREPARATION OFN-[(R)-3-DECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-HEXANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT. (COMPOUND (I), R₁═R₃═N—C₉H₁₉CO, R₂═N—C₅H₁₁CO,X═Y═O, N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

In the same manner as described in Example 16 and cognate steps, N-[(R)-3-decanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-hexanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-α-D-glucopyranosyl]-L-serinetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranoside,L-serine benzyl ester, and (R)-3-hexa- and decanoyloxytetradecanoicacids: mp 157-158° C.; IR (film) 3306, 2955, 2924, 2853, 1734, 1657,1545, 1466, 1378, 1245, 1170, 1081, 954, 842, 722 cm⁻¹; ¹H NMR(CDCl₃—CD₃ D) δ 0.88 (m, 18H), 1.15-1.7 (mH), 2.2-2.75 (m, 12H), 3.06(q, 6H, J=7.2 Hz), 3.36 (d, 1H, J=9.8 Hz), 3.43-3.63 (mH), 3.68-3.95 (m,4H), 4.13-4.27 (m, 2H), 4.54 (d, 1H, J=8.3 Hz), 4.6 (m, 1H), 5.08-5.27(m, 4H); ¹³C NMR (CDCl₃) δ 176.9, 173.4, 173.2, 172.8, 172.2, 169.5,169.1, 101.4, 74.8, 71.1, 70.9, 70.8, 69.3, 53.2, 51.6, 46.1, 41.8,41.4, 41.0, 39.2, 34.5, 34.4, 34.3, 34.1, 34.0, 32.0, 31.4, 29.8, 29.6,29.4, 29.3, 25.6, 25.3, 25.2, 25.1, 24.8, 22.7, 22.4, 14.1, 14.0, 8.7.

Anal. Calcd. for C₈₃H₁₅₈N₃O₁₉P.H₂O: C, 64.27;H, 10.40; N, 2.71; P, 2.00.Found: C, 64.09;H, 10.31; N, 2.70; P, 2.06.

EXAMPLE 37 B37 PREPARATION OFN-[(R)-3-HEXANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT. (COMPOUND (I), R₁═R₂═N—C₉H₁₉CO R₃═N—C₅H₁₁CO,X═Y═O, N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

In the same manner as described in Example 16 and cognate steps,N-[(R)-3-hexanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-α-D-glucopyranosyl]-L-serinetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranoside,L-serine benzyl ester, and (R)-3-hexa- and decanoyloxytetradecanoicacids: mp 156-157° C. dec; IR (film) 3306, 2956, 2923, 2852, 1732, 1659,1545, 1466, 1378, 1246, 1173, 1081, 958, 847, 722 cm⁻¹; ¹H NMR(CDCl₃—CD₃OD) δ 0.88 (m, 18H), 1.0-1.7 (mH), 2.2-2.75 (m, 12H), 2.9-3.3(mH), 3.06 (q, overlaps preceding multiplet, J=7.2 Hz), 3.36 (d, 1H,J=9.6 Hz), 3.43-3.63 (mH), 3.63-3.95 (m, 4H), 4.21 (m, 2H), 4.53 (d, 1H,J=8.0 Hz), 4.6 (br s, 1H), 5.06-5.28 (m, 4H); 13C NMR (CDCl₃) δ 176.6,173.6, 173.3, 172.8, 172.1, 169.6, 169.2, 101.5, 74.8, 70.9, 70.9, 69.4,60.5, 53.2, 51.5, 46.1, 41.9, 41.5, 41.1, 39.2, 34.6, 34.5, 34.4, 34.1,31.9, 31.3, 29.8, 29.7, 29.6, 29.5, 29.4, 29.3, 25.6, 25.3, 25.2, 24.7,22.7, 22.4, 14.1, 14.0, 11.1, 8.7.

Anal. Calcd. for C₈₃H₁₅₈N₃O₁₉P.H₂O: C, 64.27;H, 10.40; N, 2.71. Found:C, 64.29;H, 10.30; N, 2.61.

EXAMPLE 38 B38 PREPARATION OFN-[(R)-3-HEXANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-HEXANOYLOXYTETPADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT. (COMPOUND(I), R₁═N—C₉H₁₉CO, R₂═R₃═N—C₅H₁₁CO,X═Y═O, N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

In the same manner as described in Example 16 and cognate steps,N-[(R)-3-hexanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-hexanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-α-D-glucopyranosyl]-L-serinetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranoside,L-serine benzyl ester, and (R)-3-hexa- and decanoyloxytetradecanoicacids: mp 152-153° C. dec; IR (film) 3307, 2956, 2924, 2853, 1734, 1658,1544, 1466, 1378, 1316, 1245, 1173, 1081, 955, 843, 722 cm⁻¹; ¹H NMR(CDCl₃—CD₃OD) δ 0.88 (m, 18H), 1.15-1.7 (mH), 2.2-2.75 (m, 12H), 3.06(q, 6H, J=7.2 Hz), 3.28-3.55 (mH), 3.67-3.97 (m, 4H), 4.13-4.27 (m, 2H),4.55 (d, 2H, J=7.2 Hz), 4.60 (m, 1H), 5.08-5.28 (m,4H), 7.11 (d, 1H,J=8.7 Hz), 7.42 (d, 1H, J=8.0 Hz); ¹³C NMR (CDCl₃) δ 176.9, 173.5,173.2, 172.8, 172.2, 169.5, 169.1, 101.4, 74.8, 71.1, 70.9, 70.8, 69.3,60.5, 53.2, 51.5, 46.1, 41.8, 41.4, 41.1, 39.2, 34.5, 34.3, 34.2, 34.1,34.0, 31.9, 31.7, 31.4, 31.3, 29.8, 29.6, 29.4, 29.3, 25.6, 25.3, 25.2,24.7, 22.7, 22.4, 14.1, 14.0, 11.1, 8.7.

Anal. Calcd. for C₇₉H₁₅₀N₃O₁₉P: C, 64.24;H, 10.24; N, 2.85. Found: C,64.06;H, 10.35; N, 2.88.

EXAMPLE 39 B39 PREPARATION OFN-[(R)-3-HEXANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYETRADECANOYLAMINO]-3-O-[(R)-3-HEXANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT. (COMPOUND (I), R₁═R₃═N—C₅H₁₁CO, R₂═N—C₉H₁₉CO,X═Y═O, N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

In the same manner as described in Example 16 and cognate steps,N-[(R)-3-hexanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylaminol-3-O-[(R)-3-hexanoyloxytetradecanoyl]-α-D-glucopyranosyl]-L-serinetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranoside,L-serine benzyl ester, and (R)-3-hexa- and decanoyloxytetradecanoicacids: mp 151-152° C. dec; IR (film) 3308, 2956, 2924, 2854, 1732, 1660,1544, 1466, 1378, 1317, 1246, 1173, 1081, 957, 843, 722 cm⁻¹; ¹H NMR(CDCl₃—CD₃OD) δ 0.88 (m, 18H), 1.0-1.7 (mH), 2.18-2.72 (m, 12H), 3.06(q, 6H, J=7.4 Hz), 3.23-3.51 (mH), 3.66-3.98 (m, 4H), 4.12-4.28 (m, 2H),4.55 (d, 1H, J=7.4 Hz), 4.60 (m, 1H), 5.05-5.28 (m, 4H), 7.10 (d, 1H,J=8.2 Hz), 7.43 (d, 1H, J=8.5 Hz); ¹³C NMR (CDCl₃) δ 176.9, 173.6,173.2, 172.7, 172.2, 169.5, 169.0, 101.5, 75.0, 74.8, 71.2, 70.9, 70.8,69.2, 60.5, 53.1, 51.5, 46.1, 41.8, 41.5, 41.1, 39.1, 34.6, 34.5, 34.2,34.0, 32.0, 31.4, 31.3, 29.8, 29.7, 29.6, 29.4, 29.3, 25.6, 25.3, 25.1,24.8, 24.7, 22.7, 22.5, 22.4, 14.1, 14.0, 11.1, 8.7.

Anal. Calcd. for C₇₉H₁₅₀N₃O₁₉P.H₂O: C, 63.47;H, 10.25; N, 2.81. Found:C, 63.63;H, 10.35; N, 2.84.

EXAMPLE 40 B40 PREPARATION OFN-[(R)-3-DECANOYLOXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-HEXANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-HEXANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYL]-L-SERINETRETHYLAMMONIUM SALT. (COMPOUND (I), R₁═R₃═N—C₅H₁₁CO, R₂═N—C₉H₁₉CO,X═Y═O, N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

In the same manner as described in Example 16 and cognate steps,N-[(R)-3-decanoyloxytetradecanoyl]-0-[2-deoxy-4-O-phosphono-2-[(R)-3-hexanoyloxytetradecanoylaamino]-3-O-[(R)-3-hexanoyloxytetradecanoyl]-α-D-glucopyranosyl]-L-serinetriethylammonium salt was prepared from 2-(trimethylsilyl)ethyl2-deoxy-4,6-O-isopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranoside,L-serine benzyl ester, and (R)-3-hexa- and decanoyloxytetradecanoicacids: mp 158-159° C.; IR (film) 3308, 2956, 2924, 2854, 1734, 1659,1545, 1466, 1378, 1316, 1245, 1173, 1081, 956, 844, 722 cm⁻¹; ¹H NMR(CDCl₃—CD₃OD) δ 0.8-1.0 (m, 18H), 1.15-1.73 (mH), 2.18-2.72 (m, 12 H),3.06 (q, 6H, J=7.4 Hz), 3.35 (d, 1H, J=10 Hz), 3.47-3.67 (mH), 3.68-3.97(m, 4H), 4.1-4.3 (m, 2H), 4.54 (d, 1H, J=8.0 Hz), 4.61 (m, 1H),5.07-5.28 (m, 4H); ¹³C NMR (CDCl₃) δ 176.9, 173.5, 173.2, 172.8, 172.2,169.6, 169.1, 101.5, 75.0, 74.8, 71.2, 70.9, 70.8, 69.2, 60.5, 53.2,51.4, 46.1, 41.9, 41.5, 41.0, 39.2, 34.5, 34.2, 34.0, 31.9, 31.4, 29.8,29.6, 29.4, 29.2, 25.6, 25.3, 25.1, 25.0, 24.8, 24.7, 22.7, 22.5, 22.4,14.1, 14.0,11.1, 8.7.

Anal. Calcd. for C₇₉H₁₅₀N₃O₁₉P.H₂O: C, 63.47;H, 10.25; N, 2.81; P, 2.07.Found: C, 63.43;H, 10.22; N, 2.83; P, 2.13.

EXAMPLE 41 B41 PREPARATION OF3-HYDROXY-(R)-2-[(R)-3-DECANOYLOXYETRADECANOYLAMINO]PROPYL2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-HEXANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSIDETRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₃═N—C₉H₁₁CO, R₂═N—C₅H₁₁CO,X═Y═O, N=M=Q=0, R₄═R₅═R₇═R₉═H, R₆═OH, P=1 R₈═PO₃H₂).

In the same manner as described in Example 6 and cognate steps,3-hydroxy-(R)-2-[(R)-3-decanoyloxytetradecanoylamino]propyl2-deoxy-4-O-phosphono-2-[(R)-3-hexanoyloxytetradecanoylamino]-3-O-[(R)-3-decanoyloxytetradecanoyl]-α-D-glucopyranosidetriethylammonium salt was prepared fromN-(2,2,2-trichloroethoxycarbonylamino)-1,3,4,6-tetra-O-acetyl-2-deoxy-α-D-glucopyranoside,(S)-2-amino-3-benzyloxy-1-propanol, and (R)-3-hexa- anddecanoyloxytetradecanoic acids: mp 151-153° C.; IR (film) 3287, 2956,2923, 2853, 1732, 1643, 1552, 1466, 1378, 1318, 1147, 1176, 1108, 1082,1052, 856, 722 cm⁻¹; ¹NMR (CDCl₃—CD₃OD) δ 0.88 (t, 18H, J=6.9 Hz),1.0-1.72 (mH), 2.17-2.71 (m, 12H), 2.9-3.3 (mH), 3.08 (q, overlapspreceding multiplet, J=7.2 Hz), 3.31 (d, 1H, J=9.6 Hz), 3.5-4.02 (m,8H), 4.20 (d, 1H, J=9.5 Hz), 4.60 (d, 1H, J=8.0 Hz), 5.05-5.25 (m, 4H);¹³C NMR (CDCl₃) δ 173.7, 173.5, 173.4, 170.6, 170.1, 101.1, 75.5, 73.0,71.6, 71.3, 70.8, 70.5, 68.2, 61.4, 60.7, 54.8, 50.5, 45.8, 41.4, 39.4,34.6, 34.5, 34.2, 31.9, 31.4, 29.8, 29.7, 29.5, 29.4, 29.3, 25.4, 25.1,22.7, 22.4, 14.1, 14.0, 8.6.

Anal. Calcd. for C83H160N₃O₁₈P.H₂O: C, 64.84;H, 10.62; N, 2.55. Found:C, 65.01;H, 10.50; N, 2.55.

EXAMPLE 42 B42 PREPARATION OF5-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-DECANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYLOXY]-(S)-4-[(R)-3-DECANOYLOXYTETRADECANOYLAMINO]PENTANOICACID TRIETHYLAMMONIUM SALT (COMPOUND (I), R₁═R₂═R₃═N—C₉H₁₉CO, X═Y═O,N=M=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, P=2, R₅═PO₃H₂).

(1) In the same manner described in Example 13-(5), benzyl(S)-4-(t-butyloxycarbonylamino)-5-hydroxypentanoate (0.338 g, 0.954mmol) and the compound prepared in Example 15-(4) (1.15 g, 0.954 mmol)were coupled in the presence of AgOTf (1.22 g, 4.77 mmol) to give 0.70 g(50%) of benzyl(S)-4-(t-butyloxycarbonylamino)-5-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosyloxy)pentanoate:¹H NMR (CDCl₃) δ 0.88 (m, 6H), 1.0-2.05 (m, 55H), 2.12-2.5 (m, 6H),3.28-3.90 (m, 5H), 4.26 (dd, 1H, J=4.5, 11.5 Hz), 4.38 (d, 1H, J=11.5Hz), 4.57-4.98 (m, 5H), 5.11 (s, 2H), 5.18 (m, 1H), 5.49 (t, 1H, J=9Hz), 5.78 (d, 1H, J=7.7 Hz), 7.04-7.45 (m, 15H).

(2) A solution of the compound prepared in (1) above (0.67 g, 0.45 mmol)in CH₂Cl₂ (5 mL) was cooled to 0° C., treated dropwise with TFA (70 FL),and stirred for 3 h at room temperature. The reaction mixture wasdiluted with CH₂Cl₂ (15 mL), washed with saturated aqueous NaHCO₃ anddried (Na₂SO₄). (R)-3-Decanoyloxytetradecanoic acid (0.20 g, 0.50 mmol)and EDC Mel (0.15 g, 0.5 mmol) were added and the resulting mixture wasstirred for 16 h at room temperature. The reaction mixture was filteredthrough a pad of Celite® and concentrated. The crude product obtainedwas purified by flash chromatography on silica gel (gradient elution,15→30% EtOAc-hexanes) to give 0.36 g (45%) of benzyl(S)-4-[(R)-3-decanoyloxytetradecanoylamino]-5-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosyloxy]pentanoate:¹H NMR (CDCl₃) δ 0.89 (m, 12H), 1.0-1.978 (mH), 2.12-2.5 (m, 10H),3.45-3.65 (m, 2H), 3.79 (dd, 2H, J=3.8, 10 Hz), 4.06 (m, 1H), 4.27 (dd,1H, J=4.9, 12 Hz), 4.35 (d, 1H, J=12 Hz), 4.6-4.8 (m, 3H), 4.83 (d, 1H,J=8.3 Hz), 5.10 (s, 2H), 5.17 (m, 2H), 5.48 (t, 1H, J=10 Hz), 5.79 (d,1H, J=7.7 Hz), 6.05, (d, 1H, J=8.8 Hz), 7.07-7.42 (m, 15H).

(3) In the same manner as described for the preparation of compound B14from the compound prepared in Example 15-(5),5-[2-deoxy-4-O-phosphono-2-[(R)-3-decanoyloxytetradecanoylaminol-3-O-[(R)-3-decanoyloxytetradecanoyl]-α-D-glucopyranosyloxy]-(S)-4-[(R)-3-decanoyloxytetradecanoylamino]pentanoicacid triethylammonium salt was prepared from the compound prepared in(2) above: mp 184-188° C; IR (film) 3284, 2955, 2919, 2848, 1730, 1654,1548, 1459, 1374, 1259, 1165, 1081, 1032, 800 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD)δ 0.88 (t, 18H, J=7 Hz), 1.0-2.0 (mH), 2.18-2.75 (m, 12H), 3.08 (q, 6H,J=7.4 Hz), 3.33-4.42 (mH), 4.44 (d, 1 H, J=8.5 Hz), 5.02-5.31 (m, 4H),7.54 (d, 1H, J=8 Hz), ),7.61 (d, 1H, J=7 Hz); ¹³C NMR (CDCl₃) δ 176.8,173.6, 173.3, 170.8, 170.2, 101.1, 75.1, 73.7, 71.7, 71.1, 70.8, 70.1,60.8, 54.1, 48.8, 45.9, 41.4, 41.2, 39.4, 34.5, 34.4, 34.1, 31.9, 31.3,29.8, 29.7, 29.6, 29.5, 29.4, 26.9, 25.5, 25.3, 25.1, 22.7, 14.1, 8.6.

Negative FAB-MS calcd for [M-H]⁻ 1514.0889, found 1514.0816.

EXAMPLE 43 B43 PREPARATION OFN-[(R)-3-HYDROXYTETRADECANOYL]-O-[2-DEOXY-4-O-PHOSPHONO-2-[(R)-3-DODECANOYLOXYTETRADECANOYLAMINO]-3-O-[(R)-3-TETRADECANOYLOXYTETRADECANOYL]-α-D-GLUCOPYRANOSYL]-L-SERINETRIETHYLAMMONIUM SALT. (COMPOUND (I), R₁═N-C₁₃H₂₇CO, R₂═N—C₁₁H₂₃CO,R₃═H, X═Y═O, N=M=P=Q=0, R₄═R₅═R₇═R₉═H, R₆═CO₂H, R₈═PO₃H₂).

(1) In the same manner described in Example 13-(5),N-allyloxycarbonyl-L-serine benzyl ester (0.225 g, 0.806 mmol) and thecompound prepared in Example 22-(2) (1.104 g, 0.886 mmol) were coupledin the presence of AgOTf (0.828 g, 3.22 mmol) to give 1.01 g (83%) ofN-allyloxycarbonyl-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosyl]-L-serinebenzyl ester: ¹H NMR (CDCl₃) δ 0.88 (m, 6H), 1.0-1.7 (m, 42H), 1.78 (s,3H), 1.86 (s, 3H), 2.12-2.48 (m, 4H), 3.26 (m, 1H), 3.66 (m, 1H),3.80(dd, 1H, J=3, 10 Hz),4.19-4.38(m,4H),4.48-4.85(m,6H),4.98 (d, 1H,J=7.7 Hz), 5.08-5.38 (m, 5H), 5.49 (m, 1H), 5.60-5.75 (m, 2H), 5.82-6.0(m, 1H), 7.06-7.42 (m, 15H).

(2) A solution of the compound prepared in (1) above (1.01 g, 0.68 mmol)and diethyl malonate (1.50 g, 9.48 mmol) in THF was degassed with argon(1 h), treated with tetrakis(triphenylphosphine)palladium(0) (0.10 g,0.09 mmol), and stirred overnight at room temperature. The reactionmixture was filtered through a pad of silica with 2% MeOH—CHCl₃ and thefiltrate concentrated. A solution of the crude amine obtained in CH₂Cl₂(20 mL) was treated with (R)-3-hydroxytetradecanoic acid (0.18 g, 0.75mmol) and EDCMel (0.66 g, 1.02 mmol), stirred overnight at roomtemperature, and then concentrated. The crude product obtained waspurified by flash chromatography on silica gel (gradient elution, 30→40%EtOAc-hexanes) to give 0.506 g (46%) ofN-[(R)-3-hydroxytetradecanoyl]-O-[2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-α-D-glucopyranosyl]-L-serinebenzyl ester: ¹H NMR (CDCl₃) δ 0.88 (m, 9H), 1.0-1.7 (m, 62H),1.79(s,3H), 1.87 (s,3H),2.19(t,2H, J=7 Hz),2.3-2.5 (m, 4H),3.1 (brs, 1H), 3.55 (q, 1H, J=9 Hz), 4.0-4.43 (m, 5H), 4.56-4.85 (m, 4H), 5.13-5.32(m, 4H), 6.59 (d, 1H, J=7.4 Hz), 6.83 (br s, 1H), 7.17-7.41 (m, 15H).

(3) In the same manner as described for the preparation of compound B12from the compound prepared in Example 13-(5),N-[(R)-3-hydroxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3-dodecanoyloxytetradecanoylamino]-3-O-[(R)-3-tetradecanoyloxytetradecanoyl]-α-D-glucopyranosyl]-L-serinetriethylammonium salt was prepared from the compound prepared in (2)above: mp 170-173° C. dec; R (film) 3313, 2955, 2923, 2853, 1734, 1662,1655, 1558, 1541, 1467, 1458, 1376, 1248, 1166, 1108, 1078, 1049, 953,942, 842 cm⁻¹; ¹H NMR (CDCl₃—CD₃OD) δ 0.88 (m, 18H), 1.15-1.7 (mH),2.2-2.75(m, 12H), 3.06 (q, 6H, J=7.2 Hz), 3.3-3.63 (mH), 3.66-3.98 (m,4H), 4.1-4.3 (m, 2H), 4.54 (d, 1H, J=8.0 Hz), 4.6 (m, 1H), 5.05-5.27 (m,4H), 7.15 (d, 1H, J=8.7 Hz), 7.46 (d, 1H, J=8.2 Hz); ¹³C NMR (CDCl₃) δ177.0, 173.3, 172.8, 172.3, 171.9, 169.2, 101.2, 74.9, 74.8, 74.3, 70.8,70.6, 69.3, 68.4, 59.9, 53.1, 51.5, 42.5, 41.5, 39.2, 37.1, 34.6, 34.4,34.3, 34.1, 32.0, 29.8, 29.4, 25.6, 25.2, 22.7, 22.5, 14.5, 8.7.

Anal. Calcd. for C₈₃H₁₆₀N₃O₁₈P.2H₂O: C, 64.10;H, 10.63; N, 2.70; P,1.99. Found: C, 64.28;H, 10.42; N, 2.70; P, 1.84.

TEST EXAMPLE 1 STIMULATION OF ANTI-TETANUS TOXOID ANTIBODY PRODUCTION

The AGPs of the subject invention enhanced antibody production topurified tetanus toxoid in a murine model. Ten mg of each AGP sample wasadded to 1 ml of an oil-lecithin mixture containing squalene oil plus12% lecithin. The mixtures were heated in a 56° C. water bath andsonicated to achieve clear solutions. Fifty (50) μof each solution wasemulsified by vortexing in 2 ml of sterile, pre-warned 0.1% Tween 80saline containing 1.0 μg tetanus toxoid antigen/ml. Preparations werevortexed again just prior to administration to mice. Female C57BL/6 xDBA/2 F₁ mice (8 per group) were treated with 0.2 ml of the appropriatepreparation distributed as a 0.1 ml subcutaneous injection into eachflank. The final mouse dosage of the tetanus toxoid and AGP compoundswas 0.2 μg and 50 μg, respectively. Control mice received tetanus toxoidin vehicle (oil-Tween saline). All mice were treated on day 0 followedby a second immunization on day 21. Fourteen days following the secondimmunization mice were bled and sera were isolated by centrifugation.

Serum samples from each mouse were evaluated for anti-tetanus toxoidantibodies by enzyme immunoassay (EIA) analysis using tetanus toxoidcoated rnicrotiter plates. Anti-tetanus antibody titers were evaluatedfor IgM, total Ig, as well as, IgG₁, IgG_(2a) and IgG_(2b) isotypes.Each serum sample was diluted 2-fold for eleven dilutions starting withan initial serum dilution of 1:200. Results are shown in Tables 2-4.

TABLE 2 Anti-tetanus toxoid antibody titers of treated mice. Total IgGIgG₁ IgG_(2a) IgG_(2b) IgM Material T/C* Titer T/C Titer T/C Titer T/CTiter T/C Titer B11 3.6 23,200 1.86 400,000 2.06 10,450 0.93 26,800 4.757,600 B2 3.84 24.800 2.16 464,000 4.28 21,700 1.57 45,200 4.50 7,200 B13.97 25,600 3.42 736,000 3.78 19,200 2.45 70,400 2.38 3,800 B25 8.9357,600 2.68 576,000 1.67 8,500 3.28 94,400 2.0 3,200 B21 4.71 30,4002.23 480,000 5.83 29,600 6.07 174,400 5.50 8,800 B15 18.85 121,600 4.17896,000 6.80 34,500 2.79 80,256 4.0 6,400 Vehicle 6,450 215,000 5,07528,750 1,600 *T/C Ratio = Experimental Test Titer ÷ Vehicle ControlTiter.

TABLE 3 Anti-tetanus toxoid antibody titers of treated mice. MaterialT/C* IgM T/C IgG_(2a) T/C IgG_(2b) B12 3.1 4800 139.4 2370 149 9840 B161.6 2560 66.8 1135 104 6880 B13 3.9 6080 220 3740 >208 >13,760  B11 3.35120 347 5900 127.3 8400 Vehicle — 1760 —  25 —  98 *T/C Ratio =Experimental Test Titers ÷ Vehicle Control Titers

TABLE 4 Anti-tetanus toxoid antibody titers of treated mice. Total IgIgM IgG₁ IgG_(2a) IgG_(2b) Material T/C Titer T/C Titer T/C Titer T/CTiter T/C Titer B26 10.5 2,490 1.1 600 16.9 25,200 29.3 440 42.6 2,260B15 144.5 34,400 2.7 1,520 118.3 176,000 259.3 3,890 603.8 32,000 B2260.0 19,050 0.8 440 18.4 27,400 345.8 5,187 59.6 3,160 B28 228.6 54,5003.7 2,080 92.5 137,600 664.7 9,970 519.2 27,520 Vehicle 238 560 1,488 1553 *T/C Ratio = Experimental Test Titers ÷ Vehicle Control Titer.

Compounds of the subject invention showed a dose response whenadministered with tetanus toxoid. BFD1 (C57B1/6 X DBA/2) female mice (8per group) were immunized with 0.2 ml of emulsions containing AGP+0.2 μgof tetanus toxoid. A second immunization was administered 21 days postprimary immunization. Each mouse was bled 21 days after the secondinjection. The results are shown in Tables 5 and 6.

TABLE 5 Dose response of AGPs in mice immunized with tetanus toxoid.Total Ig IgM IgG₁ IgG_(2a) IgG_(2b) T/C T/C T/C T/C T/C Material Ratio*Titer Ratio Titer Ratio Titer Ratio Titer Ratio Titer B15 50 μg 3.37,000 13.4 37,600 4.1 26,300 150.0 11,225 3.2 2500 B15 25 μg 5.8 12,4002.1 6,000 4.5 28,800 52.0 3900 7.0 5400 B15 10 μg 5.3 11,450 1.4 4,0005.5 35,100 33.8 2538 9.9 7650 B27 50 μg 3.2 6,800 4.0 11,200 1.6 10,40012.0 900 11.6 9,000 Vehicle 2150 2800 6350 75 775 *T/C Ratio =Experimental Test Titer ÷ Vehicle Control Titer.

TABLE 6 Dose response of AGPs in mice immunized with tetanus toxoid. IgMTotal Ig IgG₁ IgG_(2a) IgG_(2b) Material T/C* Titer T/C Titer T/C TiterT/C Titer T/C Titer B12 50 μg 5.43 869 368.55 47,543 141.22 259,429 nd499.35 12,983 B12 25 μg 3.14 503 403.98 52,114 145.21 266,743 16.86 354196.92 5,120 B12 10 μg 3.71 594 248.06 32,000 81.12 149,029 6.81 143181.12 4,709 B12 5 μg 3.43 549 489.92 63,200 84.11 154,514 34.14 717352.54 9,166 B12 1 μg 1.71 274 326.02 42,057 90.08 165,486 73.71 1,548175.81 4,571 B15 50 μg 3.14 503 233.88 30,171 90.08 165,486 50.05 1,051235.62 6,126 B15 25 μg 2.29 366 181.91 23,467 106.14 194,971 10.43 219158.23 4,114 B15 10 μg 2.86 457 170.10 21,943 39.07 71,771 2.57 54 84.382,194 B15 5 μg 1.71 274 248.06 32,000 103.15 189,486 3.00 63 210.885,483 B15 1 μg 1.57 251 166.56 21,486 72.04 132,343 7.62 160 114.272,971 Vehicle 160 129 1837 21 26 *T/C = Experimental Test Titer ÷Vehicle Control Titer. nd—not done

TEST EXAMPLE 2 STIMULATION OF ANTIOVALBUMIN ANTIBODY PRODUCTION

BDF1 female mice (8 per group) were immunized with 0.2 ml of emulsionscontaining 50 μg of the AGPs+50 μg of ovalbumin. A second immunizationwas administered 21 days post primary. Each mouse was bled 14 days afterthe second injection. Antibody titers of immunized mice showing totalIgG and IgM as well as titers for the subgroups of IgG including IgG₁,IgG_(2a) and IgG_(2b) are given in Table 7.

TABLE 7 Adjuvant activity in BDF1 mice immunized with ovalbumin. TotaIIg IgM Material T/C* Titer T/C Titer B11 0.7 150 1.3 250 B2 2.5 563 0.9175 B1 0.5 119 0.8 150 B25 1.9 438 0.8 150 B21 0.5 113 1.3 250 B15 4.1925 2.3 438 B27 0.6 138 1.6 300 Vehicle — 225 — 188 IgG1 IgG2a IgG2bMaterial T/C* Titer T/C Titer T/C Titer B11 1.6 2650 1.7 550 1.6 375 B25.0 8300 2.5 825 2.3 550 B1 0.5 763 0.2 56 0.8 188 B25 5.2 8500 0.5 1635.0 1188 B21 0.6 0.1 25 0.8 200 B15 0.6 950 0.3 113 16.7 3963 B27 0.81275 0.1 38 0.5 113 Vehicle — 1650 — 325 — 238 *T/C Ratio = ExperimentalTest Titer ÷ Vehicle Control Titer

The AGP compounds of the subject invention when administered to awarm-blooded animal with the antigen ovalbumin stimulates the productionof antibody to that antigen.

TEST EXAMPLE 3 GENERATION OF A PROTECTIVE IMMUNE RESPONSE TO INFECTIOUSINFLUENZA

Mice vaccinated with formalin-inactivated influenza and the AGPcompounds of the subject invention mounted a protective immune responseto an influenza challenge as well as produced antibody to that antigen.Animals were vaccinated with the antigen and AGP compounds in variouscarriers. The degree of protection was determined by challenging themice with intranasal (IN) administration of approximately 10 LD₅₀infectious influenza A/HK/68. Mortality was assessed for 21 daysfollowing the challenge. The number of mice surviving the challenge doseis a direct assessment of the efficacy of the vaccine. For theexperiments provided this data does not necessarily correlate with theamount of antibody produced.

1) Vaccines were formulated in 0.2% triethanolamine (TEoA)/watersolution containing: 1 hemagglutinating unit (HAU) offormalin-inactivated influenza AIHK/68 (FI-Flu), and 50 μg of AGP exceptthe vehicle control vaccines which contained no AGP. ICR mice (10/group)were vaccinated 1 time only. The vaccines were administered bysubcutaneous (SQ) injection of 0.1 ml/site at 2 distinct sites near theinguinal lymph nodes for a total of 0.2 ml of vaccine/mouse. Mice (only5 mice/group) were bled from the orbital plexus 14 days following thevaccination. Sera was harvested and frozen at −20° C. until used forenzyme-linked immunosorbent assay (ELISA). All mice were challenged 30days post vaccination by intranasal (IN) administration of approximately10 LD₅₀ infectious influenza A/HK/68. Mortality was assessed for 21 daysfollowing the challenge. Anti-influenza antibody titers obtained fromvaccinations with TEoA formulations and corresponding survival rates ofmice vaccinated with this formulation are shown in Table 8.

TABLE 8 Anti-influenza antibody titers and survival rates of treatedmice. Titer⁻¹ Material Total IgG Percent Survival Nonimmune <100 0Vehicle <100 0 B9 6400 44 B10 1600 40 B7 200 33 B3 1600 33 B14 6400 44B15 6400 50

2) Vaccines were formulated in 2% Squalene solution containing: 1hemagglutinating unit (HAU) of formalin-inactivated influenza A/HK/68(FI-Flu), and 25 μg of AGP except the saline and vehicle controlvaccines which contained no AGP. BALB/c mice (10/group) were vaccinated1 time only. The vaccines were administered by subcutaneous (SQ)injection of 0.1 ml/site at 2 distinct sites near the inguinal lymphnodes for a total of 0.2 ml of vaccine/mouse. Mice (only 5 mice/group)were bled from the orbital plexus 14 days following the vaccination.Sera was harvested and frozen at −20° C. until used for enzyme-linkedimmunosorbent assay (ELISA). All mice were challenged 35 days postvaccination by intranasal (IN) administration of approximately 10 LD₅₀infectious influenza A/HK/68. Mortality was assessed for 21 daysfollowing the challenge. Anti-influenza antibody titers obtained fromvaccinations with the squalene formulations as well as correspondingsurvival rates of vaccinated animals are shown in Table 9.

TABLE 9 Anti-influenza antibody titers and survival rates of treatedmice. Titer⁻¹ Percent Material Total IgG IgG1 IgG2a IgG2b SurvivalNonimmune <100 <100 <100 <100 0 Saline 800 100 800 100 62.5 Vehicle 16001600 1600 1600 100 B25 3200 1600 6400 1600 100 B15 1600 3200 3200 400100 B9 1600 1600 3200 800 87.5 B10 400 400 400 400 62.5 B3 3200 32006400 800 87.5 B6 800 800 400 1600 75 B14 3200 6400 3200 6400 87.5 B28800 400 400 100 50

3) The antibody titers and survival rate of vaccinated mice werecompared after a primary then a secondary vaccination. Vaccines wereformulated in 0.2% TEoA/water solution containing: 1 hemagglutinatingunit of formalin-inactivated influenza A/HK/68, 25 μg AGP, except thevehicle control vaccine which contained no AGP. ICR mice (20/group) wereadministered vaccines by subcutaneous injection of 0.1 ml/site at 2distinct sites near the inguinal lymph nodes for a total of 0.2 ml ofvaccine/mouse. Each group was split into 2 subgroups 35 days after theprimary vaccination. One of each subgroup was challenged at this time,the remaining subgroups received a secondary vaccination. Mice (only5/subgroup) were bled from the orbital plexus 14 days followingvaccination (primary or secondary). Sera was 3. harvested and frozen at−20° C. until used for ELISA. Mice were challenged 35 post primary, orsecondary, vaccination by intranasal administration of approximately 10LID50, or 40 LD50, infectious influenza A/EK/68, respectively. Mortalitywas assessed for 21 days following the challenge. Anti-influenzaantibody titers and survival rates of mice post primary and postsecondary vaccination are shown in Table 10. Antibody titers as well assurvival rates of mice vaccinated a second time were higher.

TABLE 10 Antibody titers and survival rates of treated mice. IgG Titer-1Percent Survival Material post 1° post 2° post 1° post 2° Nonimmune 200100 0 0 Vehicle 800 102,400 20 40 B9 6400 12,800 80 50 B10 1600 25,60060 90 B7 3200 >102,400 60 60 B4 800 25,600 50 70 B3 3200 102,400 70 60B5 1600 >102,400 60 90 B6 1600 102,400 80 70 B14 800 51,200 33 70

TEST EXAMPLE 4 THE EFFECT OF FATTY ACID CHAIN LENGTH ON ADJUVANTICITY

The effect of the length of fatty acid chains R₁-R₃ on activity wastested. Vaccines were formulated in 0.2% TEoA/water solution containing:1 hemagglutinating unit of formalin-inactivated influenza A/HK168, and25 μg of AGP, except the vehicle control vaccines which contained noAGP. ICR mice (10/group) were vaccinated 1 time only. The vaccines wereadministered by subcutaneous injection of 0.1 ml/site at 2 distinctsites near the inguinal lymph nodes for a total of 0.2 ml ofvaccine/mouse. Mice (only 5 mice/group) were bled from the orbitalplexus 14 days following the vaccination. Sera was harvested and frozenat −20° C. until used for ELISA. All mice were challenged 35 postvaccination by intranasal administration of approximately 10 LD₅₀infectious influenza A/HK/68. Mortality was assessed for 21 daysfollowing the challenge. The length of the fatty acid chain appears tomildly affect biological activity. Results are shown in Tables 11 and12.

TABLE 11 Antibody titers and survival rates of treated mice. Titer⁻¹Total Percent Material Chain Length IgG IgG1 IgG2a IgG2b SurvivalNonimmune — 200 100 100 800  0 Vehicle — 200 100 100 200 11 B18  7 800800 800 400 20 B17  8 6400 3200 3200 1600 40 B16  9 800 1600 100 800 40B15 10 3200 200 3200 6400 70 B14 10 800 1600 100 400 30 B13 11 1600 800400 800 50 B12 12 200 200 100 200  0 B11 14 1600 200 1600 400 30

TABLE 12 Antibody titers and survival rates of treated mice. Titer⁻¹Chain Total Percent Material Length IgG IgG1 IgG2a IgG2b SurvivalNonimmune — 100 100 50 800  0 Vehicle — 100 200 50 100 30 B8  7 64003200 400 1600 80 B7  9 3200 3200 100 1600 70 B5 10 800 200 50 400 44 B411 3200 400 100 1600 60 B3 12 1600 1600 50 800  0 B1 14 12,800 6400 160015600 40

TEST EXAMPLE 5 THE EFFECT OF VARIATIONS IN THE CARBON CHAIN LENGTHBETWEEN THE HETEROATOM X AND THE AGLYCON NITROGEN ATOM ON ADJUVANTICITY

The length of the carbon chain between X and the aglycon nitrogen atomwas extended progressively by a single atom. The effect of lengtheningthe chain between these two components on adjuvanticity was explored.Vaccines were formulated in 0.2% TEoA/water solution containing: Ihemagglutinating unit of formalin-inactivated influenza A/HK/68, and 25μg of AGP, except the vehicle control vaccines which contained no AGP.ICR mice (10/group) were vaccinated 1 time only. The vaccines wereadministered by subcutaneous injection of 0.1 ml/site at 2 distinctsites near the inguinal lymph nodes for a total of 0.2 ml ofvaccine/mouse. Mice (only 5 mice/group) were bled from the orbitalplexus 14 days following the vaccination. Sera was harvested and frozenat −20° C. until used for ELISA. All mice were challenged 35 days postvaccination by intranasal administration of approximately 10 LD₅₀infectious influenza A/HK/68. Mortality was assessed for 21 daysfollowing the challenge. Adjuvant activity appears to lessen as thelength of the carbon chain between the heteroatom X and aglycon nitrogenatom increases. However, depending upon the residues attached to thiscarbon chain the biologic and metabolic stability of the molecules maybe affected. Results are shown in Tables 13.

TABLE 13 Antibody titers and survival rates of treated mice. Titer⁻¹Carbon Total Percent Material Chain IgG IgG1 IgG2a IgG2b SurvivalNonimmune — <50 <50 <50 <50  0 Vehicle — 200 200 50 200 25 B19 2 12,800100 800 6400 50 B21 3 6400 800 100 1600 40 B22 4 3200 100 3200 200 40

TEST EXAMPLE 6 CYTOKINE INDUCTION BY THE AGP COMPOUNDS

The AGP compounds of the subject invention induced cytokines in humanwhole blood ex vivo culture assays. AGP compounds were solubilized in10% EtOH-water and diluted to various concentrations. Fifty μl of eachdilution were added to 450 μl of whole human blood. Controls weretreated with culture media (RPMI). The reaction mixture was incubated at37° C. for 4 hr with constant mixing on a rotator. Sterile PBS (1.5 ml)was added to the reaction mixture, the cells were centrifuged and thesupernatents removed for cytokine testing. The concentration of TNF-αand IL-1 βin each supernatent was determined using immunoassay ELISAkits from R&D Systems. Results from these studies are shown in Tables14-19.

TABLE 14 Stimtilation of cytokine secretion in an ex vivo assay. DosageTNF-α IL-1β Material (μg) (pg/ml) (pg/ml) B26 20 498.90 33.25 10 254.9425.34 5 75.62 9.89 1 38.85 3.90 B2 20 1338.42 155.07 10 817.67 114.41 5235.32 34.72 1 105.52 14.53 RPMI — 2 0

TABLE 15 Stimulation of cytokines in an ex vivo assay. Dosage TNF-αIL-1β Material (ng/ml) (pg/ml) (pg/ml) B16 10,000   291 55 5000 277 531000 155 39 B13 10,000   775 THTC* 5000 716 187 1000 740 177 B9 10,000  449 96 5000 247 84 1000 145 53 B10 10,000   207 43 5000 127 61 1000 7317 B7 10,000   83 16 5000 57 14 1000 26 6 RPMI — 2 *THTC-To high toCount

TABLE 16 Stimulation of cytokines in an ex vivo assay. Dosage TNF-αIL-1β Material (ng/ml) (pg/ml) (pg/ml) B4 10,000   432 213 5000 205 1641000 94 70 B3 10,000   567 269 5000 390 342 1000 189 204 B5 10,000   16979 5000 143 162 1000 43 36 B6 10,000   94 52 5000 59 29 1000 30 13 B1410,000   249 91 5000 120 71 1000 56 46 RPMI — 2 0

TABLE 17 Stimulation of cytokine secretion in an ex vivo assay. DosageTNF-α IL-1β Material (ng/ml) (pg/ml) (pg/ml) B11 10,000   181 62.3 5000139 61.7 1000 115 54.5  500 125 55.8  100 127 59.8 B13 10,000   583 2825000 592 390 1000 478 327  500 411 352  100 302 261 B15 10,000   320 1535000 280 126 1000 209 94.4  500 183 104  100 133 51.6 B16 10,000 12141.0 5000 114 34.0 1000 72 19.5  500 55 17.1 B14 10,000   114 24.6 500087 19.0 1000 51 10.0  500 49 19.9 RPMI — 2 0

TABLE 18 Stimulation of cytokine secretion in an ex vivo assay. DosageTNF-α IL-1β Material (ng/ml) (pg/ml) (pg/ml) B2 10,000   100 22.2 500075 14.0 1000 38 9.0  500 28 8.3  100 6.1 3.5 B1 10,000   20 10.0 5000 115.5 1000 2.8 4.0  500 1.1 0  100 0 0 B7 10,000   61 14.7 5000 44 8.31000 30 4.3  500 27 3.8  100 10 5.1 B4 10,000   232 66.9 5000 173 66.51000 130 32.0  500 116 19.3  100 89 65.2 B3 10,000   433 151.9 5000 316200.4 1000 229 75.1  500 212 67.9  100 130 35.9 B5 10,000   142 24.15000 99 23.0 1000 96 10.5  500 59 16.9  100 33 5.4 RPMI — 2 0

TABLE 19 Stimulation of cytokine secretion in an ex vivo assay. DosageTNF-α IL-1β Material (ng/ml) (pg/ml) (pg/ml) B17 10,000   2.8 0 5000 2.20 1000 2.6 0.2 B8 10,000   2.8 0 5000 0.7 0.5 1000 1.5 0.1 B22 10,000  287 17 5000 11 1.9 1000 2.2 0.1 B28 10,000   198 13 5000 197 13 1000 1398 B12 10,000   1017 135 5000 957 153 1000 863 175 RPMI — 3.9 0

TEST EXAMPLE 7 STIMULATION OF A CYTOTOXIC T-LYMPHOCYTE RESPONSE

The induction of a cytotoxic T-lymphocyte response after administrationof the AGP compounds of the subject invention and a protein antigen wasdetected by a cytotoxicity assay. Groups of C57BIJ6 mice were given aprimary immunization subcutaneously (inguinal region) with 25 μgovalbumin (OVA) formulated in AGP preparations. The injected volume was200 μl. Twenty-one days later three mice per experimental group werekilled and spleens removed and pooled as single cell suspensions andcounted.

Spleen cells (75×10⁶ cells in 3-4 ml media) from the experimental groupswere placed in a 25 cm² T-flask. Next, 1.0 ml of irradiated (20,000rads) E.G7 (OVA) cells at 5×10⁶/ml were added to the flask. The volumewas brought to 10 ml. The cultures were maintained by placing theT-flasks upright in a 37° C., 5% CO₂ incubator for four days. On day 4the surviving cells were recovered from the flasks, washed 1X in freshmedia resuspended in 5.0 ml, and counted.

Recovered effector cells were adjusted to 5×10⁶ viable cells/ml and 100μl volumes were diluted serially in triplicate in wells of 96 wellround-bottom plates (Corning 25850) using 100 μl/well of media as adiluent. Next, 100 μl volumes of ⁵¹Cr-labelled (see below) targets [E.G7(OVA)-an ovalbumin gene transfected EL-4 cell line] at 1×10⁵ cells/mlwere added to the wells. Spontaneous release (SR) wells contained 100 μlof targets and 100 μl of media. Maximal release (MR) wells contained 100μl of targets and 100 μl detergent (2% Tween 20). Effector/target (EIT)ratios were 50:1, 25:1, 12.5:1. The plates were centrifuged at 400×g andincubated at 37° C., 5% CO₂ for 4 hr. After the incubation the wellsupernatants were collected using a Skatron Supernatant CollectionSystem.

Percent specific lysis=$100 \times \left\lbrack \frac{\left( {{{Exp}.\quad {Release}} - {SR}} \right)}{\left( {{MR} - {SR}} \right)} \right\rbrack$

Target cells, E.G7 (OVA), were labelled with ⁵¹Cr (sodium chromate) asfollows. In a total volume of 1.0 ml were mixed 5×10⁶ target cells and250 μCi ⁵¹Cr in 15 ml conical tube. The cell suspensions was incubatedin a 37° C. water bath for 90 min., with gentle mixing every 15 min.After incubation the labelled cells were washed 3× by centrifugation anddecanting with 15 ml volumes of media. After the third centrifugationthe cells were resuspended in 10 ml of fresh media and allowed to standat room temperature for 30 min. and then centrifuged. The cells werefinally resuspended in media at 1×10⁵ cells/ml.

Mice immunized according to the procedure above with the AGPs of thesubject invention displayed a cytotoxic T-lymphocyte response to the OVAantigen as shown in Table 20.

TABLE 20 Cytotoxic T-lymphocyte response of treated cells. %Cytotoxicity E:T Material 50:1 25:1 12.5:1 B11 14 8 5 B12 13 7 4 B13 2815 10 B15 58 49 30 B16 42 29 20 B17 39 26 15 B18 36 20 15 B14 45 36 25B28 28 15 9 B27 17 9 5 B1 34 24 15 B3 65 54 42 B4 72 66 60 B5 28 18 11B7 57 44 29 B8 36 20 15 B10 65 56 38 B9 65 55 36 B6 54 41 37 B2 21 12 6B25 65 55 43 B26 14 8 4 B22 58 42 31 B21 38 26 15 B19 59 42 33* B20 3625 13 B29 16 9 5 B31 19 11 7 B35 9 5 2 B36 13 7 4 B37 12 8 6 B38 38 2516 B39 33 21 13 B40 20 12 8 B43 19 12 6 Vehicle Control <10

TEST EXAMPLE 8 GENERATION OF SERUM AND MUCOSAL ANTIBODY TITERS TOTETANUS-TOXOID

The AGPs of the subject invention elicited both a serum and mucosalimmune response to purified tetanus toxoid when administeredintranasally. Groups of BALB/c mice were given a primary immunization(1°) intranasally with 10 μg tetanus toxoid (TT)+20 μg AGP formulated inan aqueous formulation (AF) in a volume of 20 μl. A secondaryimmunization (2°) was given 14 days later and a tertiary immunization(3°) identical in composition to the first and second was administered14 days later. Mice were bled on day 21 (day 7 post 2°) and day 38 (day10 post 3°) and day 48 (day 20 post 3°). Vaginal wash/fecal extractsamples were taken on day 7 post 2° and day 7 post 3°. Serum and washsamples were assayed for anti-TT antibody by standard ELISA methods.Results of these assays are shown in Tables 21 and 22 below.

The aqueous formulation comprises the AGPs of the subject invention andone or more surfactants. Surfactants useful in an aqueous compositioninclude glycodeoxycholate, deoxycholate, sphingomyelin, sphingosine,phosphatidylcholine, 1,2-Dimyristoyl-sn-glycero-3-phosphoethanolamine,L-α-phosphatidylethanolamine, and1,2-Dipalmitoyl-sn-glycero-3-phosphocholine, or a mixture thereof. Theaqueous formulation used in this example comprises the surfactant 1,2dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and was prepared asfollows: briefly; a 4 mg/ml solution of DPPC was prepared in ethanol. Analiquot of the ethanol solution is added to the dried AGPs and swirledgently to wet the AGP. The ethanol is removed by blowing a stream offiltered nitrogen gently over the vial. Water for Injection is added andthe suspension is sonicated 10 min. at 60° C. until clear. The resultingaqueous formulation contains approximately 118 μg/ml DPPC, has particlesof around 70 nm and was filter sterilized.

TABLE 21 Anti-tetanus toxoid antibody titers in treated mice.Anti-Tetanus Toxoid Titer⁻¹ Vaginal Wash Fecal Extract IgG IgA IgG IgAMaterial 2° 3° 2° 3° 2° 3° 2° 3° B25 800 6400 6400 6400 50 200 3200 6400B15 400 800 6400 6400 50 100 6400 12,800 B19 200 400 1600 3200 25 253200 6400 B4 1600 400 1600 6400 25 50 3200 12,800 B5 3200 800 3200 320050 100 3200 6400 B3 1600 1600 6400 6400 50 100 3200 6400 B22 400 800 8003200 25 50 1600 6400 PBS <25 <25 <25 <25 <25 <25 <25 <25 Normal <25 <25<25 <25 <25 <25 <25 <25 Sera

TABLE 22 Serum anti-tetanus toxoid antibody titers in treated animals.Anti-Tetanus Toxoid Titer⁻¹ Serum Pools IgG₁ IgG_(2a) IgA d21 d38 d48d21 d38 d48 d21 d38 d48 B25 1M* 8M 8M 512K 4M 4M 12.8K 102.4K 102.4K B152M 8M 8M 512K 1M 2M 12.8K 51.2K 25.6K B19 2M 4M 4M 64K# 256K 128K 6.4K25.6K 12.8K B4 1M 8M 8M 1M 2M 2M 25.6K 102.4K 102.4K B5 2M 8M 8M 512K 2M2M 25.6K 102.4K 102.4K B3 512K 4M 8M 512K 2M 2M 12.8K 51.2K 51.2K B22 1M2M 4M 64K 256K 256K 6.4K 25.6K 25.6K PBS 1,000 16K 16K 1,000 1,000 1,000200 200 200 C 200 200 200 100 100 100 200 200 200 *M = 10⁶, #K = 10³, C= normal sera

Intranasal administration of TT formulated in AGP-AF induced both anantigen specific humoral immune response (Table 22 ) and a mucosalimmune response (Table 21) to that antigen.

TEST EXAMPLE 9 STIMULATION OF AN IMMUNE RESPONSE TO HEPATITIS B SURFACEANTIGEN BY INTRANASAL ADMINISTRATION

Mice administered hepatitis B surface antigen (HBsAg) intranasally withthe compounds of the subject invention produced serum IgG and IgA titersto that antigen. Secretory IgA was detected in vaginal washes and theinduction of a cytotoxic T-lymphocyte response was detected by acytotoxicity assay.

Groups of BALB/c mice were given a primary immunization (1°)intranasally with 2.5 μg HBsAg+10 μg AGP-AF in a volume of 20 μl. AGP-AFwas prepared as in TEST EXAMPLE 8. Twenty-one days later mice were givena secondary immunization (2°) of 7.5 μg HBSAG+10 μg AGP-AF intranasallyin 20 μl volume. A tertiary immunization (3°) identical in compositionto the secondary immunization was administered 28 days after thesecondary immunization. Assays were conducted to detect cytotoxicT-lymphocyte activity at 16 days post secondary immunization (d16 post2°) and 8 days post tertiary immunization (d8 post 3°). Serum andmucosal antibody titers were assessed at 22 days post secondaryimmunization (d22 post 2°) and 21 days post tertiary immunization (d21post 3°). Antibody assays were conducted by standard ELISA methods.Cytotoxicity assays were conducted as described in TEST EXAMPLE 7.Results from this experiment are shown in Tables 23-26.

TABLE 23 Cytotoxic T-lymphocyte response of treated cells. %Cytotoxicity (d16, post 20) E/T Material 50:1 25:1 12.5:1 6.25:1 B25 3620 13 9 B15 13 5 4 4 B19 26 20 11 9 B4 28 17 9 7 B3 43 26 17 11 B5 43 3020 11 B22 33 21 15 8 Vehicle 3 2 0 0 Normal 3 3 0 0 Cells

TABLE 24 Cytotoxic T-lymphocyte response of treated cells. %Cytotoxicity (d8, post 3°) E/T Material 50:1 25:1 12.5:1 6.25:1 B25 3019 13 8 B15 56 42 25 16 B19 71 54 33 24 B4 23 15 9 5 B3 54 45 32 20 B544 30 19 12 B22 22 13 7 5 Vehicle 5 2 1 1 Normal 7 5 3 3 Cells

TABLE 25 Anti-hepatitis antibody titers in treated mice. Anti HBsAgTiter⁻¹* Material IgG₁ IgG_(2a) IgA B25 256K# 500K 3,200 B15 256K 500K6,400 B19 500K 64K 1,600 B4 500K 1000K 6,400 B3 1000K 500K 6,400 B5 256K500K 3,200 B22 256K 64K 1,600 Vehicle <2K <2K <200 *day 22 post 2°, #K =10³

TABLE 26 Anti-hepatitis antibody titers in treated mice. Anti HBsAgTiter−1* Material IgG₁ IgG_(2a) IgA B25 1000K# 1000K 25,600 B15 2000K2000K 25,600 B19 2000K 500K 12,800 B4 1000K 2000K 25,600 B3 1000K 1000K25,600 B5 500K 1000K 12,800 B22 500K 500K 12,800 Vehicle <2K <2K <200 *day 21 post 3°, #K = 10³

Groups of BALB/c mice were immunized with 2.5 μg HBsAg +10 μg AGP-AFintranasally and boosted intranasally with 7.5 μg HBsAg+10 μg AGP-AF 21days later. Vaginal samples were collected 10 days after the boosterimmunization and assayed for anti-HBsAg antibody. Results of this assayare shown in Table 27.

TABLE 27 Vaginal Wash Anti-HBsAg Titer⁻¹ Material IgG IgA B25 100 800B15 50 3200 B19 >50 400 B4 1600 6400 B3 800 1600 B5 1600 1600 B22 100800 Vehicle <50 <50

The intranasal administration of HBsAg with the compounds of the subjectinvention stimulated both a humoral and cellular immune response to thatantigen. Intranasal immunization with the antigen formulated in AGP-AFinduced a cytotoxic T-lymphocyte response (Table 23-24) and antigenspecific humoral (Table 25 and 26) and mucosal (Table 27) immuneresponses.

TEST EXAMPLE 10 GENERATION OF A PROTECTIVE IMMUNE RESPONSE TO INFLUENZA

Mice immunized intranasally with FLUSHEELD influenza vaccine containinghemagglutinin antigen and the AGPs of the subject invention producedboth IgG and IgA which were recovered in vaginal washes. Immunized micewere also protected from subsequent influenza challenge.

ICR mice were immunized three times at 21 day intervals intranasallywith FLUSHIELD influenza vaccine (Wyeth-Lederle) containing 0.3 μghemagglutinin antigen (HA)+10 μg AGP-AF or recombinant E. coli heatlabile enterotoxin (LT). AGP-AF was prepared as in TEST EXAMPLE 8. LTwas solubilized in saline at 1 μg/ml. Vaginal washes were collected 14days after the second and third immunization. Serum samples werecollected 14 days after the third immunization. Mice were challengedwith 10 LD₅₀ (lethal dose 50) of infectious influenza A/HK/68thirty-five days after the final immunization and monitored formortality. Tables 28 and 29 show the results of assays conducted bystandard ELISA methods to detect anti-influenza antibody titers invaginal washes and sera.

TABLE 28 Vaginal Wash Samples IgA IgG Percent Material SecondaryTertiary Secondary Tertiary Protection Nonimmune <20 <20 <20 <20 22Vehicle 80 160 160 160 50 B25 1280 1280 640 2560 100 B19 320 5120 12801280 70 B3 1280 2560 1280 1280 100 B22 640 2560 320 640 75 LT 2560 25602560 640 100

TABLE 29 Serum Titers Percent Material Total IgG IgG₁ IgG_(2a) IgG_(2b)Protection Nonimmune <400 <400 <400 <400 22 Vehicle 102,400 256,00012,800 102,400 50 B25 ≧819,200 102,400 819,200 ≧819,200 100  B19 819,20051,200 102,400 819,200 70 B3 ≧819,200 51,200 819,200 ≧819,200 100  B22819,200 51,200 102,400 819,200 75 LT ≧819,200 ≧819,200 ≧819,200 ≧819,200100 

These data demonstrate that AGPs in AF when administered intranasallyact as a mucosal adjuvants causing the production of IgA at mucosalsites. Increased protection is also induced against an upper respiratorypathogen which invades through the mucosa.

TEST EXAMPLE 11 GENERATION OF IMMUNE RESPONSES FROM STABLE EMULSIONFORMULATIONS

The AGP compounds of the subject invention stimulated both humoral andcytotoxic T-lymphocyte responses when formulated in a stable emulsion(SE). AGPs were tested at 25 μg dose levels to adjuvantize Hepatitis Bsurface antigen (HBsAg) for the induction of CTL and antibody responses.BALB/c mice were immunized subcutaneously with 2.0 μg HBsAg plus 25 μgof AGP/SE on day 0 and day 21. The CTL assay was conducted as in TESTEXAMPLE 7. The AGPs were formulated in a stable emulsion (SE) and thecompositions were designated AGP-SE. Methods for preparing the stableemulsion containing 10% v/v squalene, 0.091% w/v PLURONIC-F68 blockcopolymer, 1.909% w/v egg phosphatidyl choline, 1.8% v/v glycerol, 0.05%w/v a tocopherol, 10% ammonium phosphate buffer and 78.2% v/v Water forInjection should be readily apparent to one skilled in the art. Theemulsion was homogenized to a particle size of ≧0.2 μm. Table 30 showsthe AGPs of the subject invention induced a cytotoxic T-lymphocyteresponse to HBsAg.

TABLE 30 Cytotoxic T-lymphocyte response of treated cells. %Cytotoxicity E:T Material Day 50:1 25:1 12.5:1 6.25:1 B25 d17,post 1° 2712 9 5 B19 74 48 34 24 B3 28 15 9 5 B22 42 24 17 7 Vehicle-SE 32 16 9 6d16, post 2° B25 49 28 20 13 B19 73 62 42 31 B3 81 47 32 22 B22 78 69 5839 Vehicle-SE 38 23 14 8

The results of the antibody titer to HBsAg are shown on Table 31. Serafrom bleeds taken on day 28 post 2° were titered on ELISA plates coatedwith either HBsAg or a 28 amino acid peptide (p72) which contains B-cellepitopes found in the S-antigen region, residues 110-137, of the HBsAg.

TABLE 31 Anti-HBsAg titer of treated mice. Anti-HBsAg Titer⁻¹ HBsAgp72-Peptide Material IgG₁ IgG_(2a) IgG₁ IgG_(2a) B25  2048K* 2048K 128K64K B19 1024K 1024K 64K 128K B3  512K 1024K 16K 128K B22 1024K 1024K128K 128K Vehicle SE 1024K 64K 64K 4K

AGP-SE treated mice displayed both humoral (Table 31) and cytotoxicT-lymphocyte (Table 30) responses to the hepatitis B surface antigen. Ofinterest, AGP-SE treated mice in serum displayed a vigorous IgG_(2a)specific antibody titer detected by both antigens, whereas thevehicle-SE induced only a modest IgG_(2a) response.

TEXT EXAMPLE 12 STIMULATION OF SERUM ANTIBODY TITERS

The AGP compound B31 was evaluated for its ability to enhance serumantibody titers to an influenza virus vaccine as set forth in TextExample 3. In brief, ICR mice (10/group) were administered vaccinescontaining 1 HAU of formalin-inactivated influenza A/HK/68 plus or minus25 μg RC-523 formulated in a 0.2%TEoA/water solution. The mice were,also, challenged with a lethal dose of infectious influenza virus inorder to assess protection. The results of this experiment are presentedin Table 32.

TABLE 32 Anti-influenza serum titers Material IgG IgG1 IgG2a IgG2bProtection Nonimmune 200 50 50 100 0 Vehicle 200 200 50 200 25 B31 32001600 400 1600 70

TEXT EXAMPLE 13 INDUCIBLE NITRIC OXIDE SYNTHETIC ACTIVITY

Screening of respective AGP compounds of this invention includedevaluation of inducible nitric oxide synthetase or INOS activity (NOSED₅₀), which correlates with macrophage activation, and can thus beviewed as a measure of immune stimulation. For this assay, mouseperitoneal exudates cells were harvested and the adherent cellpopulation isolated. The adherent cells were exposed to varyingconcentrations of soluble AGP compounds and the resulting induction andsecretion of nitrite measured. The NOS ED₅₀ value represents aconcentration of AGP required to stimulate half the maximum amount ofnitrite release and corresponds to the concentration required tostimulate macrophages.

The AGP compounds were also evaluated for their tendency to induce afever response in rabbits. Each compound was formulated in 10% (v/v)ethanol/water solution at 100 mg/ml, then diluted with D₅W to thedesired concentration. The material was injected at 3 ml/kg body weightinto 3 rabbits. The rise in core temperature of the rabbits wasrecorded. A compound inducing a cumulative rise of greater than or equalto 1.5 degree in the three rabbits is considered pyrogenic.

The results of these experiments are presented in Table 33.

TABLE 33 NOS PYROGENICITY ED₅₀ Total Rise ° C., 3 rabbits (nanograms/ml)2.5 10 MPD # Exp. 1 Exp 2 Exp 3 ug/kg ug/kg ug/kg B1 150 0 0.1 B2 9 0.93.6 2.5-5   B3 4 0 4.2 B4 5 0.1 3.4 B5 5 3 0.1 4.1 B6 1.8 2.1 — B7 21 04 B8 ≧3000 3.6 — B9 16 0 3.1 B10 4 0 5.8 B11 3 4.2 — 0.3-0.6 B12 0.9 2.5— B13 0.1 3 — B14 0.25 2.1 — B15 0.06 4.2 3.1 <0.06 B16 0.46 1.8 — B1732.5 2.1 3.4 B18 ≧3000 4.3 — B19 100 0 0.3 B20 0.5 1.5 0.3 4.6 B21 8 0.52 B22 51 1.7 — B23 159 0.3 0.3 B24 20 17 0.9 2.4 2.5-5   B25 0.3 0.5 0.64.2 B26 67 0.2 1.7 5 B27 1.65 1.8 3.9 B28 0.3 4.2 — B29 ≧10,000 0.2 0.7B30 4.3 — B31 ≧10,000 0.5 1.6 B32 B34 ≧10,000 3.5 2.8 B35 86 3.2 — B361.8 3.4 — B37 1.1 2.2 — B38 ≧3000 3.6 — B39 ≧3000 3.2 — B40 ≧3000 3.8 —B41 6.3 — B42 5.2 — B43 380 3.8 —

It is understood that the foregoing examples are merely illustrative ofthe present invention. Certain modifications of the compositions and/ormethods employed may be made and still achieve the objectives of theinvention. Such modifications are contemplated as within the scope ofthe claimed invention.

REFERENCES

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Ikeda, K., Asahara, T. and K. Achiwa (1993) Synthesis of BiologicallyActive N-acylated L-serine-Containing Glucosaniinide-4-PhosphateDerivatives of Lipid A, Chem. Phann. Bull 41(10): 1879-1881.

Miyajima, K., Ikeda, K. and K. Achiwa (1996) Lipid A and RelatedCompounds XXXI. Synthesis of Biologically Active N-AcylatedL-Serine-Containing D-Glucosaminide 4-Phosphate Derivatives of Lipid A,Chem. Pharm. Bull. 44(12): 2268-2273.

Shimizu, T., Akiyama, S., Masuzawa, T., Yanagihara, Y., Nakamoto, S.,Takahashi, T., Ikeda, K. and K. Achiwa (1985) Antitumor Activity andBiological Effects of Chemically Synthesized Monosaccharide Analogues ofLipid A in Mice. Chem. Pharm. Bull. 33(10): 4621-4624.

Shimizu, T., Sugiyama, K., Iwamoto, Y., Yanagihara, Y., Asahara, T.,Ikeda, K. and K. Achiwa (1994) Biological Activities of ChermicallySynthesized N-acylated Serine-linked Lipid A Analog in Mice, Int. J.Immunopharmac., 16(8): 659-665.

Shimizu, T., Iida, K., Iwamoto, Y., Yanagihara, Y., Ryoyama, K.,Asahara, T., Ikeda, K. and K. Achiwa (1995) Biological Activities andAntitumor Effects of Synthetic Lipid A Analogs Linked N-Acylated Serine,Int. J. Immunopharmac., 17(5): 425-431.

What is claimed is:
 1. An immunoeffector compound having the followingstructure:

wherein, X is selected from the group consisting of O and S at the axialor equitorial position; Y is selected from the group consisting of O andNH; n, m, p and q are integers from 0 to 6; R₁, R₂ and R₃ are the sameor different and are normal fatty acyl residues having from 1 to about20 carbon atoms and where one of R₁, R₂ or R₃ is optionally hydrogen; R₄and R₅ are the same or different and are selected from the groupconsisting of H and methyl; R₆ and R₇ are the same or different and areselected from the group consisting of H, hydroxy, alkoxy, phosphono,phosphonooxy, sulfo, sulfooxy, amino, mercapto, cyano, nitro, formyl andcarboxy, and esters and amides thereof; and R₈ and R₉ are the same ordifferent and are selected from the group consisting of phosphono and H,and at least one of R₈ and R₉ is phosphono.
 2. The compound of claim 1,wherein R₆ is carboxy.
 3. The compound of claim 2, wherein X is O; Y isO; n, m, p and q are 0; R₁, R₂ and R₃ are normal fatty acyl residueshaving 10 carbon atoms; R₄, R₅ and R₇ are H; R₈ is phosphono; R₉ is H;R₁, R₂ and R₃ are each attached to a stereogenic center having an Rconfiguration; and R₅ is attached to a stereogenic center having an Sconfiguration.
 4. The compound of claim 2, wherein X is O; Y is O; n, m,p and q are 0; R₁, R₂ and R₃ are normal fatty acyl residues having 12carbon atoms; R₄, R₅ and R₇ are H; R₈ is phosphono; R₉ is H; R₁, R₂ andR₃ are each attached to a stereogenic center having an R configuration;and R₅ is attached to a stereogenic center having an S configuration. 5.The compound of claim 2, wherein X is O; Y is O; n, m, p and q are 0;R₁, R₂ and R₃ are normnal fatty acyl residues having 10 carbon atoms;R₄, R₅ and R₇ are H; R₈ is phosphono; R₉ is H; R₁, R₂ and R₃ are eachattached to a stereogenic center having an R configuration; and R₅ isattached to a stereogenic center having an R configuration.
 6. Thecompound of claim 2, wherein X is O; Y is O; n, m, p and q are 0; R₁, R₂and R₃ are normal fatty acyl residues having 8 carbon atoms; R₄, R₅ andR₇ are H; R₈ is phosphono; R₉ is H; R₁, R₂ and R₃ are each attached to astereogenic center having an R configuration; and R₅ is attached to astereogenic center having an S configuration.
 7. The compound of claim1, wherein R₆ is H.
 8. The compound of claim 7, wherein X is O; Y is O;n is 2; m, p and q are 0; R₁, R₂ and R₃ are normal fatty acyl residueshaving 14 carbon atoms; R₄, R₅ and R₇ are H; R₈ is phosphono; R₉ is H;and R₁, R₂ and R₃ are each attached to a stereogenic center having an Rconfiguration.
 9. The compound of claim 7, wherein X is O; Y is O; n is1, m and p are 0; q is 1; R₁, R₂ and R₃ are normal fatty acyl residueshaving 10 carbon atoms; R₄ and R₅ are H; R₇ is carboxy; R₈ is phosphono;R₉ is H; and R₁, R₂ and R₃ are each attached to a stereogenic centerhaving an R configuration.
 10. The compound of claim 7, wherein X is O;Y is O; m, n, p and q are 0; R₁, R₂ and R₃ are normal fatty acylresidues having 14 carbon atoms; R₄, R₅ and R₇ are H; R₈ is phosphono;R₉ is H; and R₁, R₂ and R₃ are each attached to a stereogenic centerhaving an R configuration.
 11. The compound of claim 7, wherein X is O;Y is O; m, n, p and q are 0; R₁, R₂ and R₃ are normal fatty acylresidues having 10 carbon atoms; R₄, R₅ and R₇ are H; R₈ is phosphono;R₉ is H; and R₁, R₂ and R₃ are each attached to a stereogenic centerhaving an R configuration.
 12. The compound of claim 7, wherein X is O;Y is O; m, p and q are 0; n is 1; R₁, R₂ and R₃ are normal fatty acylresidues having 14 carbons; R₄, R₅ and R₇ are H; R₈ is phosphono; R₉ isH; and R₁, R₂ and R₃ are each attached to a stereogenic center having anR configuration.
 13. The compound of claim 1, wherein R₆ is hydroxy. 14.The compound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is1; R₁, R₂ and R₃ are normal fatty acyl residues having 12 carbon atoms;R₄ and R₅ are H; R₇ is H; R₈ is phosphono; and R₉ is H; R₁, R₂ and R₃are each attached to a stereogenic center having an R configuration; andR₅ is attached to a stereogenic center having an S configuration. 15.The compound of claim 13, wherein X is O; Y is O; m and q are 0; n and pare 1; R₁, R₂ and R₃ are normal fatty acyl residues having 10 carbonatoms; R₄, R₅ and R₇ are H; R₈ is phosphono; R₉ is H; R₁, R₂ and R₃ areeach attached to a stereogenic center having an R configuration; and R₅is attached to a stereogenic center having an S configuration.
 16. Thecompound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 2;R₁, R₂ and R₃ are normal fatty acyl residues having 10 carbon atoms; R₄,R₅ and R₇ are H; R₈ is phosphono; R₉ is H; R₁, R₂ and R₃ are eachattached to a stereogenic center having an R configuration; and R₅ isattached to a stereogenic center having an S configuration.
 17. Thecompound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 1;R₁, R₂ and R₃ are normal fatty acyl residues having 14 carbon atoms; R₄,R₅ and R₇ are H; R₈ is phosphono; R₉ is H; R₁, R₂ and R₃ are eachattached to a stereogenic center having an R configuration; and R₅ isattached to a stereogenic center having an R configuration.
 18. Thecompound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 1;R₁, R₂ and R₃ are normal fatty acyl residues having 14 carbon atoms;R₄,R₅ and R₇ are H; R₈ is phosphono; R₉ is H; R₁, R₂ and R₃ are eachattached to a stereogenic center having an R configuration; and R₅ isattached to a stereogenic center having an S configuration.
 19. Thecompound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 1;R₁, R₂ and R₃ are normal fatty acyl residues having 11 carbon atoms; R₄,R₅ and R₇ are H; R₈ is phosphono; R₉ is H; R₁, R₂ and R₃ are eachattached to a stereogenic center having an R configuration; and R₅ isattached to a stereogenic center having an S configuration.
 20. Thecompound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 1;R₁, R₂ and R₃ are normal fatty acyl residues having 10 carbon atoms; R₄,R₅ and R₇ are H; R₈ is phosphono; R₉ is H; R₁, R₂ and R₃ are eachattached to a stereogenic center having an R configuration; and R₅ isattached to a stereogenic center having an S configuration.
 21. Thecompound of claim 1, wherein X is O; Y is O; m, n, p and q are 0; R₁, R₂and R₃ are normal fatty acyl residues having 10 carbon atoms; R₄ and R₅are H; R₆ is amino carbonyl; R₇ is H; R₈ is phosphono; and R₉ is H; R₁,R₂ and R₃ are each attached to a stereogenic center having an Rconfiguration; and R₅ is attached to a stereogenic center having an Sconfiguration.
 22. The compound of claim 1, wherein R₁ is hydrogen. 23.The compound of claim 1, wherein R₂ is hydrogen.
 24. The compound ofclaim 1, wherein R₃ is hydrogen.
 25. A method for enhancing the immuneresponse of a mammal comprising administering to the mammal an effectiveamount of a compound of claim
 1. 26. A vaccine composition comprising acompound of claim 1, an antigen and a suitable carrier.
 27. Apharmaceutical composition comprising a compound of claim 1 and apharmaceutically acceptable carrier.
 28. The composition of claim 27,wherein said pharmaceutically acceptable carrier is an aqueouscomposition comprising water and one or more surfactants selected fromthe group consisting of glycodeoxycholate, deoxycholate, sphingomyelin,sphingosine, phosphatidylcholine,1,2-Dimyristoyl-sn-glycero-3-phosphoethanolamine,L-α-Phosphatidylethanolamine, and1,2-Dipalmitoyl-sn-glycero-3-phosphocholine, or a mixture thereof. 29.The composition of claim 28, wherein said one or more surfactant is1,2-Dipalmitoyl-sn-glycero-3-phosphocholine.
 30. The composition ofclaim 28, wherein the molar ratio of said compound to surfactant is fromabout 10:1 to about 1:25.
 31. The composition of claim 28, wherein themolar ratio of said compound to surfactant is from about 4:1 to about1:9.
 32. The composition of claim 27, wherein said carrier is a stableemulsion comprising a metabolizable oil, one or more surfactants, anantioxidant and a component to make the emulsion isotonic.
 33. Thecomposition of claim 32, wherein said stable emulsion comprises 1-10%v/v squalene, 0.9% w/v PLURONIC-F68 block co-polymer, 1.9% w/v eggphosphatidyl choline, 1.75% v/v glycerol and 0.05% w/v a tocopherol. 34.The composition of claim 27 wherein said carrier is a suspensioncomprising aluminum hydroxide, calcium hydroxide, calcium phosphate ortyrosine adsorbate.
 35. The composition of claim 27 wherein said carrieris an aqueous solution or aqueous micellar dispersion comprisingtriethylamine or triethanolamine.
 36. The composition of claim 27wherein said carrier comprises microspheres or microparticles, and thecompound of claim 1 is within the matrix of the microspheres ormicroparticles or adsorbed thereon.